U.S. patent application number 12/485842 was filed with the patent office on 2010-06-24 for synthesis of oligomers in arrays.
This patent application is currently assigned to LIFE TECHNOLOGIES CORPORATION. Invention is credited to John H. Butler.
Application Number | 20100160185 12/485842 |
Document ID | / |
Family ID | 35732738 |
Filed Date | 2010-06-24 |
United States Patent
Application |
20100160185 |
Kind Code |
A1 |
Butler; John H. |
June 24, 2010 |
Synthesis of Oligomers in Arrays
Abstract
Systems, including apparatus and methods, for synthesis of
oligomers in arrays.
Inventors: |
Butler; John H.; (San Jose,
CA) |
Correspondence
Address: |
LIFE TECHNOLOGIES CORPORATION;C/O INTELLEVATE
P.O. BOX 52050
MINNEAPOLIS
MN
55402
US
|
Assignee: |
LIFE TECHNOLOGIES
CORPORATION
Carlsbad
CA
|
Family ID: |
35732738 |
Appl. No.: |
12/485842 |
Filed: |
June 16, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11172753 |
Jun 30, 2005 |
|
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12485842 |
|
|
|
|
60584524 |
Jun 30, 2004 |
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Current U.S.
Class: |
506/40 |
Current CPC
Class: |
B01J 2219/00418
20130101; B01J 2219/00689 20130101; B01J 2219/00725 20130101; B01J
2219/00414 20130101; B01J 19/0046 20130101; B01J 2219/00454
20130101; B01J 2219/00511 20130101; B01J 2219/00423 20130101; B01J
2219/0036 20130101; B01J 2219/005 20130101; B01J 2219/00317
20130101; B01J 2219/00596 20130101; B01J 2219/00722 20130101; B01J
2219/00497 20130101; B01J 2219/00286 20130101; B01J 2219/0059
20130101; B01J 2219/00378 20130101; B01J 2219/00585 20130101 |
Class at
Publication: |
506/40 |
International
Class: |
C40B 60/14 20060101
C40B060/14 |
Claims
1. A device for parallel synthesis of oligomers, comprising: a
porous member including an array of islands and a spacer joined to
the array of islands and separating the islands, the array of
islands being more hydrophilic than the spacer, each island
defining a plurality of pores permitting reagents to pass through
such island; a channel structure disposed adjacent the porous
member and including a plurality of reaction compartments disposed
in one-to-one correspondence with the array of islands; and a set
of particles disposed in each reaction compartment and retained
therein by the channel structure, the set of particles being
configured to support synthesis of at least one oligomer using the
reagents that pass through the corresponding island of the
array.
2. A device for directing fluids, comprising: a porous member
including an array of islands and a spacer joined to the array of
islands and separating the islands, the islands and the spacer
having different surface energies, the porous member including
opposing surfaces, each island defining a network of pores and
being configured to receive reagents and to permit passage of the
reagents between the opposing surfaces.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of U.S. application Ser.
No. 11/172,753 filed Jun. 30, 2005, which claims the benefit under
35 USC .sctn.119(e) of U.S. Provisional Application No. 60/584,524
filed Jun. 30, 2004, both of which are incorporated herein by
reference in their entirety.
INTRODUCTION
[0002] Oligomers are chemical compounds, such as oligonucleotides
or peptides, that include a covalently linked chain of individual
subunits. The identity of each individual subunit and the sequence
of the individual subunits within the chain generally define the
chemical and biological properties of each oligomer. In particular,
a small change in the chemical structure of an oligomer, such as a
single nucleotide change in an oligonucleotide, can impart quite
distinct biological properties to the oligomer. Accordingly, large
sets of oligomers can be synthesized for use in various clinical
and research applications.
SUMMARY
[0003] The present teachings provide systems, including apparatus
and methods, for synthesis of oligomers in arrays.
BRIEF DESCRIPTION OF THE DRAWINGS
[0004] FIG. 1 is a schematic view of an exemplary system for
solid-phase synthesis of oligomers in an array using a synthesis
support device having a porous member that defines an array of
addressable sites, in accordance with aspects of the present
teachings.
[0005] FIG. 2 is a schematic view of the synthesis support device
of FIG. 1, in accordance with aspects of the present teachings.
[0006] FIG. 3 is a view of another exemplary system for solid-phase
synthesis of oligomers in an array using a synthesis support device
having a porous member that defines an array of addressable sites,
in accordance with aspects of the present teachings.
[0007] FIG. 4 is a sectional view of the synthesis support device
of FIG. 3, taken generally along line 4-4 of FIG. 3, in accordance
with aspects of the present teachings.
[0008] FIG. 5 is fragmentary sectional view of an exemplary
addressable site (and reaction compartment) from the synthesis
device of FIG. 4, taken generally from the region indicated at "5"
in FIG. 4.
[0009] FIG. 6 is a series of fragmentary sectional views of
configurations of the addressable site of FIG. 5 as the site is
being addressed with a first reagent in fluid isolation from other
addressable sites of the synthesis support device, and then with a
second reagent in fluid communication with the other addressable
sites, in accordance with aspects of the present teachings.
[0010] FIG. 7 is a series of fragmentary sectional views of
configurations of the addressable site of FIG. 5 during
post-synthesis processing of oligomer populations synthesized on
solid support surfaces of the site, in accordance with aspects of
the present teachings.
[0011] FIG. 8 is a series of views of a porous member being
processed so that regions of the porous member are addressable
selectively, in accordance with aspects of the present
teachings.
[0012] FIG. 9 is a series of view of structures produced during
fabrication of a channel (well) array for assembly with the
selectively addressable porous member produced in FIG. 8 to form an
array-defining portion of a synthesis support device, in accordance
with aspects of the present teachings.
DESCRIPTION OF VARIOUS EMBODIMENTS
[0013] The present teachings provide systems, including apparatus
and methods, for synthesis of oligomers in arrays. The apparatus
can include a synthesis support device that defines an array of
addressable sites (and reaction compartments). This array of sites
can be defined by a porous member having (1) a plurality of porous
islands, and (2) a spacer of different surface energy (e.g.,
wettability) than the porous islands that separates the porous
islands. For example, the islands can be more hydrophilic (or less
hydrophobic) than the spacer. Accordingly, the islands can be
addressed in isolation with smaller volumes of fluid, such as with
selected reagents for oligomer synthesis, because the different
surface energy of the spacer can restrict lateral movement of the
fluid away from the islands. In addition, the islands can be
addressed in fluid communication with one or more common reagents
using larger volumes of fluid that can flood the islands and
thereby overcome the surface energy barrier created by the spacer.
Methods of synthesizing oligomers using porous members also are
disclosed.
[0014] Oligomers can be synthesized on reaction surfaces that
overlap and/or are spaced from the porous islands. For example, the
reaction surfaces can be provided by reaction compartments defined
by the porous islands and/or in reaction compartments disposed
adjacent the porous islands, such as on adjoining channel walls
and/or particles, among others. Accordingly, each oligomer can be
synthesized on two or more distinct reaction surfaces that can
provide separate populations of the oligomer for the same or
different purposes, such as structural analysis (e.g., sequence
verification and/or quality control) and/or experimental/diagnostic
use, among others. Therefore, the systems of the present teachings
can offer (1) greater flexibility of solid-phase oligomer synthesis
and/or (2) synthesis of a larger amount of each oligomer in a
smaller area and/or with a smaller amount of reagents than systems
that synthesize oligomers on a nonporous planar surface, in
microplate wells, and/or in separate columns.
[0015] FIG. 1 shows an exemplary system 20 for synthesizing
oligomers in an array. System 20 can include a synthesis support
device or platform 22 that defines an array of addressable sites
and reaction compartments for solid-phase oligomer synthesis.
System 20 also can include a reagent dispenser 24, a flow
controller 26, and/or a processor 28, among others.
[0016] Reagent dispenser 24 can be configured to dispense reagents
selectively to individual sites of device 22 and/or to dispense one
or more common reagents nonselectively to many or all of the
addressable sites of the support device.
[0017] Flow controller 26 can exert a pressure periodically or
continuously on the synthesis support device, to move reagents into
and/or through reaction compartments. Accordingly, the flow
controller can include a pump, a centrifuge, and/or pressurized
gas, among others.
[0018] Processor 28 can be a data processor or controller
configured to control and coordinate any suitable aspects of system
20. For example, processor 28 can control positioning of the
reagent dispenser, can select reagents and volumes thereof to be
dispensed, and/or can operate valves and/or pumps that effect
dispensing of the selected reagents and volumes. Alternatively, or
in addition, processor 28 can control operation of flow controller
26, such as selecting the times at which pressure is exerted on the
synthesis support device, thereby controlling reaction (or reagent
contact) times during oligomer synthesis.
[0019] FIG. 2 shows a schematic representation of synthesis support
device 22. Support device 22 can include a porous member 42,
reaction compartments 44, and/or an adjoining chamber(s) or
receiver compartment(s) 46, among others. Porous member 42 can
define an array of addressable regions or sites, generally as an
array of porous islands 48 separated by a spacer of different
surface energy than the islands. Reagents can be received in each
of the porous islands for placement in reaction compartments 44
having reaction surfaces 50 for supporting solid-phase oligomer
synthesis. Alternatively, or in addition, the reaction compartments
can be used to conduct liquid-phase oligomer synthesis, during
which oligomer intermediates are not coupled to a solid support
surface.
[0020] Reaction compartments 44 and/or reaction surfaces 50 can be
disposed in one-to-one correspondence with the porous islands.
Accordingly, the reaction compartments and reaction surfaces can
define arrays aligned with, adjacent, and/or overlapping the array
of porous islands 48 included in the porous member. For example,
reaction surfaces 50 can be in reaction compartments included in
the porous islands and/or disposed adjacent the porous islands. In
some examples, at least a portion of each reaction surface can be
included in a reaction compartment configured as a channel (a
permeable well) adjoining each porous island. The channel can have
a reaction surface defined by a wall of the channel and/or by a
support matrix, such as particles, disposed in the channel, among
others. The channel can be configured to receive a reagent from a
corresponding porous island, to temporarily retain the reagent, and
then to permit the reagent to be removed from the channel. The
reagent can be removed by fluid flow out of the channel into
adjoining chamber 46, which can serve as a waste reservoir. Fluid
flow can be created by a pressure created by a flow controller in
fluid communication with the adjoining chamber.
[0021] Further aspects of the present teachings are described in
the following sections, including (I) synthesis support devices,
(II) reagent dispensers and reagents, (III) reagent removal
mechanisms, and (IV) examples.
I. Synthesis Support Devices
[0022] The synthesis systems of the present teachings can include
one or more synthesis support devices. A synthesis support device
generally includes any device having an array of reaction surfaces
that are individually addressable with selected reagents and
capable of supporting oligomers during their synthesis. Support
devices can include an array portion with a porous member and
reaction compartments. The support device also can include a body
portion or frame holding the array portion and at least partially
defining an adjoining chamber(s). Further aspects of support
devices are included in the following subsections, including (A)
porous members, (B) reaction compartments, and (C) adjoining
chambers.
[0023] A. Porous Members
[0024] A porous member, also termed a filtration device, generally
includes any structure having a plurality of pores permitting
passage of fluid between opposing surfaces of the structure.
[0025] The porous member can have any suitable configuration and
can be formed of any suitable material(s). The porous member can be
one-piece (unitary), or it can be formed of or two or more distinct
pieces. In some examples, the porous member can be generally
planar, with its thickness substantially less than its length and
width. The thickness of the porous member can be selected, for
example, according to a desired mechanical strength, ease of
fabrication, a desired fluid capacity per unit area (according to
pore size/density), and/or the like. In some examples, the porous
member can be nonplanar. The porous member can have any suitable
size and can be large enough to define an array of any suitable
size. The porous member can be formed of a material that is a
conductor (including a metal or a semi-conductor, among others) or
an insulator. Exemplary materials that can substantially form the
porous member can include silicon, gallium, a metal(s), a metal
alloy(s), plastic, ceramic, glass, and/or any combination thereof,
among others.
[0026] The porous member can include a plurality of porous islands
or porous regions separated by a spacer. Each porous island or
region can be configured to permit fluid communication between
opposing surfaces of the island. Accordingly, the porous island
includes at least one pore (or opening), and generally a plurality
of pores (or openings), configured to permit such fluid
communication. The porous island can have any suitable shape,
including rectangular, circular, ovalloid, elliptical, etc.
[0027] Porous islands can be present in any suitable number and in
any suitable arrangement. The porous islands can have a regular or
irregular spacing. In exemplary embodiments, the porous islands are
disposed in a rectilinear arrangement of two or more rows and two
or more columns. However, other suitable arrangements can include
one row or one column and can include a radial array, a staggered
(e.g., hexagonal) array, an irregular array, and/or the like. In
exemplary embodiments, the porous islands (and associated reaction
compartments) can define an array of about 100 to 5,000 islands. In
some examples, the array can correspond in spacing and arrangement
to a microplate array, for example, a microplate with 96, 384, or
1536 wells, among others. In such examples, the center-to-center
spacing between islands can be about 9 mm, 4.5 mm, or 2.25 mm,
among others, and the arrangement of islands can be an 8.times.12,
16.times.24, or 32.times.48 rectangular array, among others.
[0028] The spacer can be any intervening material that at least
substantially or completely separates the porous islands. The
spacer can be joined to the porous islands, for example, formed
with the porous islands from a single piece of material.
Accordingly, the spacer can be porous and can have pores that are
similar (or different) in size and/or shape to those of the porous
islands.
[0029] The porous islands and the spacer can have different surface
energies to provide differential wettabilities or surface tensions.
These differences can be local and/or average differences, among
others. Different surface energies, as used herein, can be a
differential affinity for fluid sufficient or effective to restrict
lateral movement or spreading of a liquid from an island to the
spacer (and thus from an island to an adjacent island). For
example, the porous islands can have a higher surface energy than
the spacer, so that the porous islands are relatively hydrophilic
and the spacer is relatively hydrophobic. A polar liquid (such as
water or an aqueous solution) thus can be selectively addressed to
one (or more) of a plurality of hydrophilic islands separated by a
hydrophobic spacer. This setup is particularly suitable for
synthesis of water-soluble oligomers or polymers, such as nucleic
acids and proteins. Conversely, the porous islands can have a lower
surface energy than the spacer, so that the porous islands are
relatively hydrophobic and the spacer is relatively hydrophilic. A
nonpolar liquid (such as an organic solvent) thus can be
selectively addressed to one (or more) of a plurality of
hydrophobic islands separated by a hydrophilic spacer. This setup
is particularly suitable for synthesis of water-insoluble oligomers
or polymers. Generally, relatively higher wettabilities imply a
greater tendency for a fluid to spread on a solid surface and be
imbibed by a porous surface, and relatively lower wettabilities
imply a lesser tendency for a fluid to spread on a solid surface
and be imbibed by a porous surface. The surface energy can be a
surface energy of an exterior surface of regions of the porous
member and/or of an interior surface defined by pores. Differences
in surface energy between the islands and the spacer can be created
by differential surface modification/treatment of the islands or
the spacer and/or by forming the islands and spacer out of
different materials having different surface energies, among
others. For example, one of the islands and spacer can be treated
with a wetting agent, and/or the other of the islands and spacer
can be treated with a nonwetting or waterproofing agent, among
others.
[0030] The relative affinity between a liquid and a solid surface
can be characterized by the contact angle between the liquid in
contact with the solid surface. This angle is determined by
competition between liquid-liquid molecular forces and liquid-solid
molecular forces, and so depends in part on the particular solid
and liquid involved, as well as the smoothness and cleanliness of
the surface. Generally, the smaller the contact angle, the greater
the affinity between the liquid and the surface, and the more
easily the liquid will penetrate pores formed by the surface. In
particular, in pores penetrated by capillary action, the fluid will
rise (or extend) nearer the walls of the pore for contact angles
less than 90 degrees (with 0 degrees being totally flat or spread),
and the fluid will fall (or recede) nearer the walls for angles
greater than 90 degrees (with 180 degrees between totally rounded
up or spherical). However, the total penetration of liquid into the
pore will be determined by an interplay between contact angle,
surface tension, and fluid density, among others. Thus, for a given
liquid, the islands and the spacer can be distinguished by
different contact angles, typically less than 90 degrees for one,
and greater than 90 degrees for the other.
[0031] Pores, as used herein, are openings of any suitable diameter
and shape. The pores can be macropores or nanopores. Macropores, as
used herein, have an average diameter of equal to or greater than
about one micrometer, and nanopores have an average diameter of
less than about one micrometer. Generally, capillary action will
draw fluid more easily into small pores, and less easily into large
pores, all other things being equal. The pores can be an
interconnected set or network of pores or can follow separate paths
between opposing surfaces of the porous member. The pores can be
present at any suitable density to achieve any suitable
permeability and fluid capacity of a porous member.
[0032] Pores can be created by any suitable process. The pores can
be created mechanically (e.g., using a drill), optically (e.g.,
using a laser), chemically (e.g., by wet-etching), electrically
(e.g., by using a nonporous member as an electrode), and/or as
voids within an assembly of fibers (such as a fiber filter), among
others. In exemplary embodiments, the pores are formed by
wet-etching a silicon wafer.
[0033] B. Reaction Compartments
[0034] The synthesis support device can include a plurality of
reaction compartments in fluid communication with, overlapping
with, and/or at least substantially coextensive with the porous
islands. A reaction compartment can include any space for receiving
reagents and having a reaction surface(s) to support synthesis of
an oligomer(s) using the reagents.
[0035] The reaction compartment can be configured to hold fluid
transiently and to permit removal of the fluid. Accordingly, the
reaction compartment can be defined by and/or disposed adjacent a
porous or permeable structure. For example, the reaction
compartment can be defined by a porous island of a porous member,
with the walls of the pores being reaction surfaces of the
compartment. Alternatively, or in addition, the reaction
compartment can be or include a space disposed adjacent the porous
island.
[0036] The space adjacent the porous island can be a channel that
permits fluid flowthrough. The channel can be configured to receive
reagents at a first end of the channel and to release at least a
portion of these reagents for removal at a second end of the
channel. The first and second ends can be generally opposing.
Accordingly, the first and second ends of the channel can be
permeable, provided, for example, by (1) a porous member (such as a
porous island thereof) and (2) a permeable layer flanking the
channel. Although the permeable layer can permit fluid flow, the
permeable layer can be configured to reduce fluid flow so that
reagents are retained at least transiently in the channel to permit
chemical reactions to occur.
[0037] The channel can include or contain any suitable reaction
surfaces to support oligomer synthesis. For example, the channel
can have a wall defining a reaction surface. Alternatively, or in
addition, the channel can hold a matrix or discrete particles (such
as beads) having reaction surfaces. The particles can have any
suitable size or shape and can be formed of any suitable material,
including plastic (such as polystyrene, among others), glass (such
as controlled-pore glass (CPG)), metal, etc.
[0038] The space adjacent the porous island, in some embodiments,
can be created by a well having a nonpermeable end/bottom wall. In
these embodiments, the reagents can be received and removed from
the same region of the well.
[0039] The reaction surface can be any solid and/or persistent
surface (including a gel) to which oligomer intermediates are
connected during oligomer synthesis. The reaction surface can
provide a covalent linkage to oligomer intermediates (and
oligomers). Accordingly, the reaction surface can include a first
reactive moiety configured to react to form a covalent bond with a
second reactive moiety of an oligomer subunit or intermediate (or a
precursor thereof). Exemplary pairs of first and second (or second
and first) reactive moieties can be classified as electrophilic and
nucleophilic moieties, as presented in Table 1. Here, persistent
means that the surface remains at least substantially intact or
functional during the course of a surface-associated reaction.
TABLE-US-00001 TABLE 1 Chemically Reactive Moieties Electrophilic
Moiety Nucleophilic Moiety Resultant Covalent Linkage activated
esters amines/anilines carboxamides acyl azides amines/anilines
carboxamides acyl halides amines/anilines carboxamides acyl halides
alcohols/phenols esters acyl nitriles alcohols/phenols esters acyl
nitriles amines/anilines carboxamides aldehydes amines/anilines
imines aldehydes or ketones hydrazides hydrazones aldehydes or
ketones hydroxylamines oximes aldehydes or ketones
thiosemicarbazides thiosemicarbazones alkyl halides amines/anilines
alkyl amines alkyl halides carboxylic acids esters alkyl halides
thiols thioethers alkyl halides alcohols/phenols ethers alkyl
sulfonates thiols thioethers alkyl sulfonates carboxylic acids
esters alkyl sulfonates alcohols/phenols ethers anhydrides
alcohols/phenols esters anhydrides amines/anilines carboxamides
aryl halides thiol thiophenols aryl halides amines aryl amines
azindines thiols thioethers boronates glycols boronate esters
carboxylic acids amines/anilines carboxamides carboxylic acids
alcohols esters carboxylic acids hydrazines hydrazides
carbodiimides carboxylic acids N-acylureas or anhydrides
diazoalkanes carboxylic acids esters epoxides thiols thioethers
haloacetamides thiols thioethers halotriazines amines/anilines
ammotriazines halotriazines alcohols/phenols triazinyl ethers imido
esters amines/anilines amidines isocyanates amines/anilines ureas
isocyanates alcohols/phenols urethanes isothiocyanates
amines/anilines thioureas maleimides thiols thioethers
phosphoramidites alcohols phosphite esters silyl halides alcohols
silyl ethers sulfonate esters amines/anilines alkyl amines
sulfonate esters thiols thioethers sulfonate esters carboxylic
acids esters sulfonate esters alcohols ethers sulfonyl halides
amines/anilines sulfonamides sulfonyl halides phenols/alcohols
sulfonate esters
[0040] Alternatively, or in addition, the reaction surface can
provide a noncovalent association with oligomer intermediates (and
completed oligomers), such as binding through a specific binding
pair (antibody-antigen, receptor-ligand, enzyme-substrate,
complementary nucleotide strands, etc.). The reaction surface for
noncovalent or covalent association can be any suitable external or
internal surface(s) of the synthesis support device including the
walls of a channel, the walls of pores, the walls of a well, and/or
the exterior/interior surface of particles, among others.
[0041] A reaction compartment can include two or more distinct
reaction surfaces permitting coupled oligomers to be separated
during and/or after their synthesis. The distinct reaction surfaces
can be physically separable, that is, disposed on separable
structures. Alternatively, or in addition, the distinct reaction
surfaces can be chemically distinct so that oligomers can be
selectively removed from one or more of the reaction surfaces.
Accordingly, distinct reaction surfaces of a reaction compartment
can provide oligomer coupling that is selectively sensitive to any
suitable uncoupling treatment, such as pH, heat, light, exposure to
a particular chemical cleavage agent, etc.
[0042] C. Adjoining Chambers
[0043] A synthesis support device can have one or more chambers
adjoining the array portion of the support device. An adjoining
chamber can be substantially enclosed so that the chamber can hold
a reduced (or increased) pressure, to draw (push) fluid from the
reaction compartments to (away from) the chamber. The adjoining
chamber can be a single chamber in fluid communication with an
entire array of islands/reaction compartments, for concurrent
application of an increased or decreased pressure to the
islands/reaction compartments. Alternatively, the adjoining chamber
can be a plurality of chambers, in fluid communication with
individual islands/reaction compartments or subsets of two or more
islands/reaction compartments. Configuration of the adjoining
chamber as a plurality of chambers can permit selective removal of
fluid from a subset of the reaction compartments. In some examples,
the synthesis support device can include at least one adjoining
chamber configured to receive reagents from the reaction
compartments, thereby serving as a waste reservoir.
[0044] The adjoining chamber can be created by a chamber structure
adjoining the array portion (e.g., the porous member) of the
support device. The chamber structure can form a substantial seal
against a surface of the array portion, for example, against an
upper surface, a lower surface, and/or a perimeter of the array
portion. The chamber structure can be disposed generally above
and/or below the array portion of the synthesis support device, for
example, as a cover for the array portion and/or as a frame that
supports the array portion.
II. Reagent Dispensers and Reagents
[0045] The synthesis systems of the present teachings can include
one or more reagent dispensers configured to dispense reagents to a
synthesis support device. A reagent dispenser can include a
dispense head, reagent reservoirs, conduits, valves, and/or pumps,
among others. The reagent dispenser can dispense reagents using
contact and/or noncontact mechanisms.
[0046] Each reagent dispenser can dispense reagents to addressable
sites of the array portion from one or more dispense heads, each
having one or more dispense structures (such as dispense tips,
among others). The dispense structures of a dispense head can be
fixed and/or movable in relation to the addressable sites. The
dispense structures can fixed or movable within a dispense head. In
some embodiments, the systems described herein can include two or
more dispense heads that are movable independently. Such dispense
heads can be configured to dispense the same reagents as each other
(redundant dispense heads) or different reagents. If the same or
overlapping sets of reagents are dispensed by two or more dispense
heads, corresponding dispense structures of the dispense heads can
be connected to the same reagent reservoir or different reservoirs.
The use of two or more dispense heads (and/or the use of two or
more dispense tips per dispense head) can increase synthesis
throughput.
[0047] Reagent reservoirs disposed in fluid communication with the
dispense structures can store any suitable number of reagents. The
dispense structures can be connected in one-to one correspondence
with a set of reagent reservoirs. Alternatively, different reagent
reservoirs can be in communication with the same dispense
structure, to provide, for example, mixed and/or alternate
dispensing of reagents from the different reagent reservoirs.
[0048] The reagent dispenser can include conduits, valves, and/or a
pump to propel, guide, and/or restrict movement of reagents between
the reagent reservoirs and the dispense structures. The conduits
can define parallel paths between the reagent reservoirs and the
dispense structure. Alternatively, or in addition, the conduits can
define a branched network so that the same reagent reservoir can
connect to a plurality of dispense structures and/or so that a
plurality of reagent reservoirs can connect to the same dispense
structure. The valves (or one valve) can open and close the
conduits and can be operable manually and/or through a controller.
The open time for a valve can define the volume of reagent
dispensed to a reaction vessel. The pump (or pumps) can be any
mechanism that propels reagents from the reagent reservoirs to the
dispense structures and/or that expels reagents from the dispense
structures. The pump can exert a pressure on reagents directly or
on a compartment in fluid communication with the reagents. The pump
can act to push and/or pull reagents during dispensing (e.g., by
creating positive relative pressure within a dispense tip to push
reagents out, or by creating a negative relative pressure outside
the dispense tip to pull reagents out, respectively, among others).
Accordingly, the pump can be a positive-displacement pump (e.g., a
syringe pump, a peristaltic pump, a rotary pump, etc.), a vacuum
pump, pressurized gas, a partial vacuum, and/or the like. In some
embodiments, the gas (such as argon, among others) provided by the
pump places reagents under a more inert environment, such as by
reducing exposure to moisture, oxygen, etc. In some embodiments,
the dispenser can include nozzles configured to dispense small
volumes of some of or all of the reagents, for example, using
inkjet technology, such as a piezoelectric dispense mechanism
and/or a thermal dispense mechanism, among others.
[0049] The reagent dispenser can dispense any suitable reagents for
synthesis of oligomers. Such reagents, generally termed oligomer
reagents, can include oligomer components and ancillary
reagents.
[0050] Oligomer components generally include any chemical compounds
that are partially or completely incorporated into oligomers during
their synthesis, generally through covalent linkage. Oligomer
components can be configured so that reactive groups are protected,
exposed, and/or created relative to parent compounds, as
appropriate. An oligomer component can correspond to a portion or
all of a subunit of an oligomer, a dimer of subunits, a trimer of
subunits, etc. Exemplary oligomer components include nucleic acid
components, such as deoxyribonucleotides, ribonucleotides, peptide
nucleic acids, locked nucleic acids, or analogs, relatives,
derivatives (e.g., phosphoramidite derivatives thereof, among
others), or portions thereof. In some examples, the oligomer
components can include adenosine, cytidine, guanosine, and
thymidine phosphoramidites held in individual reservoirs, to be
addressed individually (or in combination) to reaction
compartments. Alternatively, or in addition, the oligomer
components can include nucleotide derivatives with modified bases.
Other exemplary oligomer components include amino acids, or
analogs, relatives, derivatives, or portions thereof, to form
peptides or peptide analogs (peptidomimetics). Additional exemplary
oligomer components can include carbohydrates, lipids, metalorganic
compounds, etc.
[0051] Ancillary reagents can include any other reagents that
facilitate oligomer synthesis. Such ancillary reagents can include
a solvent or fluid carrier, reagents for capping (protection of
reactive groups), deprotection, oxidation, reduction, cyclization,
washing, cleavage (uncoupling from a reaction surface), etc. In
some embodiments, the fluid carrier can be or include acetonitrile.
Alternatively, or in addition, the fluid carrier can be or include
a high-boiling point liquid (solvent), such as described in U.S.
Pat. Nos. 6,177,558 and 6,337,393 to Brennan et al., each of which
is incorporated herein by reference. In some embodiments, reagents
can be configured to perform a gas/vapor phase cleavage, as
described, for example, in U.S. Pat. No. 5,514,789 to Kempe, which
is incorporated herein by reference.
[0052] Oligomers generally include any molecule formed of two or
more covalently linked subunits. The term oligomer, as used herein,
also is intended to encompass polymers of any size or complexity.
Accordingly, an oligomer can have any suitable number of subunits,
for example, greater than ten, greater than one-hundred, or greater
than one-thousand subunits, among others. The various subunits of
an oligomer can be structurally identical (such as oligomers with a
repeated subunit), structurally related but including distinct
subunits (such as oligomers of different nucleotides or amino
acids), and/or structurally unrelated (such as oligomers including
different structural classes of subunits), as desired. Oligomers
synthesized by the systems described herein can have a predefined
size (or length), composition, and/or sequence of subunits.
However, such oligomers can be synthesized as mixtures of
oligomers, such as degenerate oligonucleotides synthesized with a
mixture of nucleotide components at one or more positions of the
oligonucleotides.
[0053] The oligomers can be used for any suitable purpose(s). For
example, nucleic acids oligomers can be used in genomics
applications, such as gene expression analysis, detection of
single-nucleotide polymorphisms, and/or high density TAQMAN assays,
among others. Accordingly, nucleic acid oligomers can be used as
probes (e.g., fluorescence in situ hybridization (FISH) probes),
primers (e.g., polymerase chain reaction (PCR) primers),
substrates, test compounds for screens, and/or reagents, among
others. Amino acid polymers similarly can be used as probes,
primers, substrates (e.g., enzyme substrates such as kinase
substrates), biological modulators, test compounds for screens,
and/or reagents, among others.
III. Reagent Removal Mechanisms
[0054] Reagents, including reacted derivatives thereof, can be
removed from reaction compartments using one or more reagent
removal mechanisms of an oligomer synthesis system. A reagent
removal mechanism can remove excess/unreacted reagent from a
reaction compartment to a waste reservoir, such as an adjoining
chamber, for example, as described above in Section I. The reagent
removal mechanism can be configured to remove reagents from the
reaction compartments of the array at substantially the same time
or at different times for subsets of the array. Alternatively, or
in addition, the reagent removal mechanism can be configured to
move reagents within the array portion of a synthesis support
device, such as movement from a porous member to an adjoining
reaction compartment, among others.
[0055] The reagent removal mechanism can be configured to push
and/or pull fluid from a reaction compartment. For example, the
reagent removal mechanism can exert a positive pressure to push the
fluid through the reaction compartment. Alternatively, or in
addition, the reagent removal mechanism can exert a negative
pressure to pull the fluid from the reaction compartment. The
pressure exerted by the reagent removal mechanism can be adjustable
and controlled by a processor to define the rate of movement of
reagents through a reaction compartment, time of contact with the
reagents, etc. In some examples, reagents can be removed by
centrifugation of the synthesis support device.
[0056] The reagent removal mechanism can be configured to operate
on one reaction compartment at a time, on a set of two or more
reaction compartments at the same time, or on all of the reaction
compartments concurrently. Accordingly, the reagent removal
mechanism can be disposed adjacent each of the reaction
compartments concurrently or can move among the reaction
compartments of a synthesis support device, for example by sliding
back and forth and/or movement along two axes, among others, to
selectively remove reagent(s) from a subset of the reaction
compartments.
IV. Examples
[0057] The following examples describe selected aspects and
embodiments of the present teachings, including an exemplary
synthesis system for synthesizing oligomers in an array, support
devices for oligomer synthesis in arrays, and methods of making and
using such support devices. These examples and the various features
and aspects thereof are included for illustration and are not
intended to define or limit the entire scope of the present
teachings.
Example 1
Exemplary System for Oligomer Synthesis in an Array
[0058] This example describes an exemplary system for oligomer
synthesis in an array; see FIG. 3.
[0059] Synthesis system 70 can include an array device 72, a
reagent dispenser 74, a flow controller 76, and a computing device
78, among others. Reagent dispenser 74 can dispense reagents to
array device 72. Flow controller 76 can be disposed in fluid
communication with the array device for continuous or periodic
removal of the dispensed reagents (or portions/derivatives thereof)
from reaction compartments of the array device. Computing device 78
can be disposed in communication with array device 72, reagent
dispenser 74, and/or flow controller 76 for operation and/or
monitoring thereof.
[0060] Array device 72 can include a chamber structure or frame 80
and an array portion 82 held by the frame. Array portion 82 can
define an array 84 of selectively addressable reaction compartments
86. Frame 80 and array portion 82 can cooperatively define a
chamber 88 in which fluid is received from the reaction
compartments. Frame 80 can be connected to tubing 90 that provides
fluid communication between flow controller 76 and chamber 88.
[0061] Reagent dispenser 74 can include a dispense head 92,
reagents reservoirs 94, and conduits 96 connecting the reagent
reservoirs to the dispense head. The dispense head 92 can include
one or more dispense tips 98 from which reagents 100 of the reagent
reservoirs 94 can be dispensed, shown at 102. Dispense head 92
and/or tips 98 can be movable (such as orthogonally), shown at 104,
to position the head or tips for selective delivery to reaction
compartments 86. Alternatively, or in addition, array device 72 can
be movable to position reaction compartments in relation to
dispense head 92 and/or tips 98.
[0062] Computing device 78 can control and coordinate operation of
system 70. For example, computing device 78 can be configured to
select a reagent reservoir(s) from which a reagent will be
dispensed, to select a position for head 92, and to operate a pump
and/or valve(s) to control the volume of the reagent that is
delivered. Computing device 78 can operate flow controller 76 to
control exposure of the reaction compartments to reagents.
Computing device 78 also or alternatively can be in communication
with one or more sensors of the system. The sensors can be
configured to sense any suitable aspects of the system, including
temperature, reagent status, pressure, dispensed volume, reaction
efficiency, etc.
Example 2
Exemplary Synthesis Support Device
[0063] This example describes an exemplary synthesis support
device, and reaction compartments and reaction surfaces thereof;
see FIGS. 4 and 5. Selected aspects of this synthesis support
device were discussed above in Example 1, particularly in the
context of FIG. 3.
[0064] Support device 72 can include an array portion 82 formed by
one or more apposed layers of material. The apposed layers can
include a porous member 122, a channel layer 124, and a permeable
retainer layer 126.
[0065] Porous member 122 can form a top layer of array portion 82.
The porous member can include a plurality of porous islands 128
separated by a spacer 130 to define array 84. The porous islands
can have any suitable arrangement to define an array of addressable
regions. The porous islands can be substantially more hydrophilic
than the spacer, for example, so that the spacer is hydrophobic and
the islands hydrophilic.
[0066] Channel layer 124 can define an intermediate or lower layer
of array portion 82. The channel layer can form an array of
channels or through-holes 132 extending between opposing surfaces
134, 136 of the channel layer. Channels 132 can correspond in
number and arrangement to the porous islands, so that channels 132
and porous islands 128 are substantially aligned and present in
one-to-one correspondence. Each channel 132 can contain one more
particles 138.
[0067] Retainer layer 126 can define a lower layer of array portion
82. The retainer layer can be configured to retain particles 138
(and fluid) in channels 132. The retainer layer also can be
permeable to permit fluid flow through this layer. The retainer
layer can be any permeable material, including a fiber filter
(e.g., formed of glass fibers, cellulose, synthetic polymer
strands, etc.), a layer of porous silicon, or the like.
[0068] Array portion 82 can be assembled from two or more layers,
such as the sandwich of three layers shown herein, and can be
supported by frame 80. Frame 80 can include a support flange 140 to
support the array portion above chamber 88. Frame 80 also can
include an opposing flange 142 that opposingly grips the array
portion with support flange 140. Opposing flange 142 can be
adjustable to restrict removal of array portion 82 and/or
separation or relative movement of the layers thereof during
oligomer synthesis. Opposing flange 142 can be removable to allow
processing of the array portion during and/or after oligomer
synthesis.
[0069] During oligomer synthesis, reagents can be addressed to each
porous island from dispense tips 98 of dispense head 92, shown at
144. Reagents can be received by porous islands 128 and can flow to
channels 132 for contact with particles 138 therein. After
reaction, the reagents (or portions/derivatives thereof) can be
moved from channel 132, through retainer layer 126 and into waste
reservoir 88, shown at 146, to join waste fluid 148.
[0070] FIG. 5 shows an individual reaction compartment 170 of array
portion 82 of the synthesis support device 72. Reaction compartment
170 can be formed by porous island 128, channel 132, particles 138,
or a combination thereof, among others.
[0071] Porous island 128 can include a plurality of pores 172,
which are shown schematically in the present illustration.
Individual pores or sets of pores 172 can extend between opposing
surfaces 174, 176 of porous member 122 to permit passage of fluid
through the island. Pores can be defined by pore walls 178 that
include pore reaction surfaces 180. Pore reaction surfaces 180 can
include a first reactive moiety 182 covalently or noncovalently
coupled to the pore walls. Accordingly, pores 172 can be configured
to create an island reaction sub-compartment 184.
[0072] Channel 132 can be configured to be defined by channel walls
186 having channel reaction surfaces 188. Channel reaction surfaces
can include a second reactive moiety 190 coupled to channel walls
186. In the present illustration, second reactive moiety 190 is
distinct from first reactive moiety 182 of the porous member. In
some examples, the same reactive moiety can be coupled to each of
the porous member and the channel layer, and/or a plurality of
different reactive moieties can be included in one or more of the
surfaces. Channel 132 can form a channel reaction sub-compartment
192 adjoining the island reaction sub-compartment 184.
[0073] Channel reaction sub-compartment 192 can contain particles
138 having particle reaction surfaces 194. Particle reaction
surfaces 194 can include a third reactive moiety 196, which can be
the same or different from the other reactive moieties. In the
present illustration, third reactive moiety 196 is different from
first and second reactive moieties 182, 190. Use of different
reactive moieties can provide support for synthesis of different
oligomers within a reaction compartment and/or selective uncoupling
of a subset of an oligomer population from a reaction
compartment.
Example 3
Placement of Reagents in Reaction Compartments
[0074] This example describes exemplary methods of addressing
reaction compartments in fluid isolation or fluid communication;
see FIG. 6. In the present sequence of configurations, first and
second reagents 202, 204 are shown being dispensed sequentially to
reaction compartment 170 with the first reagent in isolation from,
and the second reagent in communication with, other reaction
compartments of array portion 82.
[0075] First reagent 202 can be dispensed from above (and/or
adjacent) porous island 128, shown at 206 in the first
configuration. The first reagent can be dispensed in a droplet(s)
208 having a volume insufficient to spread laterally beyond the
island after contact with the porous island. Accordingly, the
placed droplet, shown at 210 in the second configuration, can be
restricted substantially to the porous island and restricted from
flowing laterally over or into spacer 130 by a difference in
surface energy of the porous island and the surrounding spacer.
First reagent 202 thus can be received in reaction compartment 170,
in fluid isolation from other reaction compartments, shown at 212
in the third configuration. The first reagent can be received by a
force or pressure exerted on the first reagent (such as a vacuum, a
positive pressure, or a centrifugal force) and/or by capillary
action.
[0076] Second reagent 204 can be dispensed to array portion 82
after the first reagent, shown at 214 after placement, in the
fourth configuration of the sequence. First reagent 202 can be
substantially removed from reaction compartment 170 before
dispensing second reagent 204, as shown in the present
illustration, or second reagent 204 can be dispensed to porous
island 128 before substantial removal of first reagent 202 from the
reaction compartment. The second reagent can be dispensed in a
fluid volume sufficient to spread laterally beyond the island, to
"flood" the upper surface of the porous member, indicated at 216 in
the fourth configuration. As a result, second reagent 204 can be
introduced into concurrent contact with a plurality of the porous
islands (and/or all of the porous islands) with a single dispensing
operation. Furthermore, the second reagent can be received
substantially concurrently in each of the reaction compartments of
the array portion 82 from above the porous islands, shown for one
of the reaction compartments at 218 in the fifth configuration. The
second reagent can be removed after being received in the reaction
compartment, shown at 220 in the sixth configuration.
Example 4
Oligomer Release
[0077] This example describes exemplary methods of releasing
oligomers from various reaction surfaces of a support device; see
FIG. 7.
[0078] Configuration 230 of the sequence shows completed oligomer
populations 232, 234, 236 connected, respectively, to support
surfaces of porous member 122, particles 138, and channel wall 186
of array portion 82. These oligomer populations can represent
structurally identical or distinct oligomer populations. A subset
of the oligomer populations, such as oligomer populations 232 and
234, can be selectively uncoupled (cleaved) from their support
surfaces by a cleavage treatment, indicated at step 238. In the
present illustration, step 238 includes a vapor phase
cleavage/deprotection, without elution of the uncoupled oligomer
populations. Accordingly, prior to cleavage, a subset of the
oligomer populations, such as oligomer populations 232, 234, can be
coupled to their support surfaces using a different association
mechanism than the remaining oligomer population(s), such as
oligomer population 236. Configuration 240 shows oligomer
populations 232, 234 uncoupled from their support surfaces but not
removed from regions adjacent these surfaces.
[0079] Porous member 122 can be separated from the channel and
retainer layers 124, 126, shown at step 242. The resultant
configurations 244, 246 each can be disposed adjacent a sample
holder 248, for example, a sample holder having wells 250 or other
receiving compartments arranged according to the array of reaction
sub-compartments of porous member 122 and channel layer 124.
[0080] A force can be applied to each of the porous member and the
channel layer, shown respectively at steps 252 and 254. The force
can be applied by a vacuum pump, pressurized gas, and/or by
centrifugation, among others. The force can elute oligomer
populations 232, 234 from their respective support structures into
wells 250, shown in configurations 256, 258. In some examples,
additional fluid, such as an aqueous buffer, can be added to one or
both reaction sub-compartments to facilitate elution of the
oligomer populations from the porous member and channel layer.
Eluted oligomers can be analyzed according to their concentration,
purity, sequence, and/or the like, for example, for quality control
purposes. Alternatively, or in addition, eluted oligomers can be
used for any suitable assay(s). The eluted oligomers produced by
steps 252, 254 can form duplicate or corresponding arrays for the
same or different purposes.
[0081] Channel layer 124 can be separated from retainer layer 126
and particles 138, shown at step 260 and represented by a partially
disassembled state in configuration 262. Particles 138 can be
discarded at this stage.
[0082] Configuration 264 shows oligomer population 236 being
cleaved from its support surface by a laser 266 and subsequently
analyzed. Oligomer population 236 can be coupled to channel layer
124 by a linker 190 that is resistant to the vapor phase cleavage
treatment of step 238. Accordingly, oligomer population 236 can
remain coupled to its support surface during the processing steps
preceding step 260. Linker 190 can be a photocleavable linker
(e.g., o-nitro benzyl, among others), so that light 268 from laser
266 can be used to cleave the oligomer in a vacuum. The cleaved
oligomer thus can be analyzed by travel through an electric field
by mass spectrometry, such as by matrix assisted laser desorption
ionization (MALDI). Accordingly, channel layer 124 can be formed of
a conductive material, such as silicon, among others.
Example 5
Fabrication of Support Devices
[0083] This example describes exemplary methods of fabricating
support devices for synthesis of oligomers in an array; see FIGS. 8
and 9.
[0084] FIG. 8 is a series of views of a porous member being
processed according to a method 270 of forming a porous member
having an array of hydrophilic islands and a hydrophobic spacer.
Hydrophilic porous member 272 can be formed from a nonporous member
by any suitable treatment, such as chemical etching of a silicon
wafer, among others, or may be rendered porous by its fabrication
(such as a fiber filter).
[0085] Step 274 can be performed by treatment of first porous
member 272 with a modifying agent to create a hydrophobic porous
member 276 from a more hydrophilic first porous member. In
exemplary embodiments, step 274 can be performed by treatment of a
silicon porous member with a fluorosilane or an alkane, among
others. In some embodiments, step 274 can be performed selectively,
such as with a mask, to create hydrophilic islands and a
hydrophobic spacer.
[0086] Step 278 can be performed next to add a first mask layer 280
to hydrophobic porous member 276. The first mask layer can be
patterned or not patterned, as shown in the present illustration.
In some examples, step 278 can include forming a layer of a
positive (or negative) photoresist on a surface of the hydrophobic
porous member, such as by spin coating, among others.
[0087] Next, step 282 can place a second mask layer 284 on first
mask layer 280 to create assembly 286. Second mask layer 284 can be
a pre-patterned mask layer, such as a quartz chromium mask layer
having optically transparent and opaque regions 288, 290,
respectively.
[0088] Step 292 can be performed next by exposure of assembly 286
to light 294. Light is permitted to pass through transparent
regions 288 and restricted from passage through opaque regions 290.
Accordingly, first mask layer 280 can be selectively exposed to the
light according to the arrangement of transparent regions 288.
[0089] The second mask layer then can be removed and the first mask
layer processed to selectively remove regions exposed (or not
exposed) to the light, shown at step 296, thereby creating
uncovered regions 298.
[0090] Step 302 can be performed next to selectively modify
uncovered regions 298 of hydrophobic porous member 276 to create a
patterned porous member 304 having porous islands 306 and a
hydrophobic spacer 308.
[0091] First mask layer 280 then can be removed, for example, by
stripping photoresist from patterned porous member 304, shown at
step 310. Porous member 304 then can be used for selective
placement of reagents and/or to support oligomer synthesis in an
array of reaction compartments.
[0092] Patterned porous member 304 can be fabricated by any
suitable variations of method 270. For example, a pre-patterned
mask layer, such as a quartz chromium mask, can be placed directly
onto hydrophobic porous member 276, without use of a first mask
layer. Next, hydrophobic porous member 276 can be selectively
ablated adjacent transparent positions of the mask by exposure to
light, such as ultraviolet light, to selectively increase the
hydrophilicity of regions of the porous member.
[0093] FIG. 9 is a series of views of structures produced by a
method 320 of fabricating a channel (permeable well) array. The
channel array can be assembled with the patterned porous member
produced by method 270 of FIG. 8 to create an array portion of a
synthesis support device. The channel array can include a substrate
322, such as a silicon substrate, that is patterned by method
320.
[0094] A first mask layer 324 can be applied to substrate 322,
shown at step 326. The first mask layer can be, for example, a
positive or negative photoresist.
[0095] A pre-patterned mask layer 328 then can be placed on first
mask layer, shown at step 330, to form a substrate assembly 332.
The pre-patterned mask layer can, for example, define a predefined
spatial pattern of permissive and restrictive light transmission,
such as with a quartz chromium mask.
[0096] Substrate assembly 332 can be exposed to light, shown at
step 334. The light can selectively photolyze regions of the first
mask layer apposed to transparent regions of pre-patterned mask
layer 328 (see method 270 of FIG. 8).
[0097] The pre-patterned mask layer 328 and photolyzed regions of
the first mask layer then can be removed, shown at step 336.
[0098] Substrate 322 then can be exposed to an etchant configured
to selectively remove the substrate at unprotected regions 338 to
create channels 340 in a channel layer 342, shown at step 344. Any
suitable etchant can be used including a chemical etchant (such as
hydrofluoric acid), anodization (such as pulse anodization), and/or
photoinduction, among others. Channels 340 can be through-holes of
any suitable shape, include cylindrical, frustoconical, etc.
Alternatively, channels 340 can be recesses having a porous
floor.
[0099] First mask layer 324 then can be removed and channels 340
modified to create derivatized channel layer 346, shown at step
348. Channel modification can include reacting surfaces of the
channels with a bis functional moiety (two or more reactive
groups), for example, to coat the channel surfaces with a reactive
moiety. Next, the reactive moiety on the channel surface can be
connected to a photocleavable linker by chemical reaction.
Alternatively, a photocleavable linker can be selected that is
directly reactive with the channel surfaces without prior
modification using the bis functional moiety. A suitable
photocleavable linker can be stable during oligomer synthesis.
[0100] Channel layer 346 then can be further modified to create
channel assembly 348, shown at step 350. In particular, a permeable
retainer layer 352 can be apposed to the channel layer to create
permeable wells 354. Optionally, particles 356 can be placed in the
permeable wells and retained therein by retainer layer 352. Porous
member 304 of method 270 (see FIG. 8) can be placed over channel
assembly 348 to form a filtration device defining an array of
selectively addressable reaction compartments.
Example 6
Further Aspects of the Present Teachings
[0101] This example suggests potential advantages of the synthesis
platform of the present teachings over other platforms. These
advantages can include one or more of the following.
[0102] (1) Scalability: the synthesis scale can be defined by the
dimensions of the well (or channel), the area of the solid
support(s) (e.g., particle surface area), and/or the concentration
of reactive moieties on the solid support(s).
[0103] (2) The number of oligomers produced per synthesis run can
be defined by the platform design (for example, the number of
reaction compartments per synthesis support device).
[0104] (3) Cycle times can be directly related to the number of
nozzles used to dispense amidites (or other oligomer reagents) and
the speed and accuracy of dispensing.
[0105] (4) Reagent use can be minimized through surface tension
localization.
[0106] (5) Substrates can be used as oligomer arrays where quality
control can be performed using fluorescence-based strategies.
[0107] (6) Arrays can be used for genomics applications.
[0108] (7) The platform can be compatible with various oligomer
chemistries, such as peptide nucleic acids, locked nucleic acids,
peptides, small molecules, and/or the like.
[0109] (8) Oligomers can be synthesized with a low cost per
oligomer.
[0110] The disclosure set forth above may encompass multiple
distinct inventions with independent utility. Although each of
these inventions has been disclosed in its preferred form(s), the
specific embodiments thereof as disclosed and illustrated herein
are not to be considered in a limiting sense, because numerous
variations are possible. The subject matter of the inventions
includes all novel and nonobvious combinations and subcombinations
of the various elements, features, functions, and/or properties
disclosed herein. The following claims particularly point out
certain combinations and subcombinations regarded as novel and
nonobvious. Inventions embodied in other combinations and
subcombinations of features, functions, elements, and/or properties
may be claimed in applications claiming priority from this or a
related application. Such claims, whether directed to a different
invention or to the same invention, and whether broader, narrower,
equal, or different in scope to the original claims, also are
regarded as included within the subject matter of the inventions of
the present disclosure.
* * * * *