U.S. patent application number 16/914486 was filed with the patent office on 2020-10-15 for storage of codes in molecularly imprinted polymers.
This patent application is currently assigned to JOHANNES KEPLER UNIVERSITAT LINZ. The applicant listed for this patent is JOHANNES KEPLER UNIVERSITAT LINZ, Klaus MOSBACH. Invention is credited to Oliver BRUGGEMANN, Klaus MOSBACH, Jacqueline WOLFSCHLUCKER.
Application Number | 20200327936 16/914486 |
Document ID | / |
Family ID | 1000005117869 |
Filed Date | 2020-10-15 |
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United States Patent
Application |
20200327936 |
Kind Code |
A1 |
MOSBACH; Klaus ; et
al. |
October 15, 2020 |
STORAGE OF CODES IN MOLECULARLY IMPRINTED POLYMERS
Abstract
Disclosed is a molecularly imprinted polymer for storing a
defined value of a numerical code, more particularly a binary code,
in the molecular imprints of said polymer, and a method for the
production of said polymer. The molecular imprinting process uses
suitable templates comprising a defined sequence of at least two
different structural units, each having a different chemical
functionality.
Inventors: |
MOSBACH; Klaus; (Zurich,
CH) ; BRUGGEMANN; Oliver; (Wilhering, AT) ;
WOLFSCHLUCKER; Jacqueline; (Zwettl an der Rodl, AT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MOSBACH; Klaus
JOHANNES KEPLER UNIVERSITAT LINZ |
Zurich
Linz |
|
CH
AT |
|
|
Assignee: |
JOHANNES KEPLER UNIVERSITAT
LINZ
Linz
AT
|
Family ID: |
1000005117869 |
Appl. No.: |
16/914486 |
Filed: |
June 29, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16096008 |
Oct 24, 2018 |
10734072 |
|
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PCT/AT2017/060111 |
Apr 27, 2017 |
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16914486 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
B01J 20/268 20130101;
C08L 101/00 20130101; G01N 24/08 20130101; G11C 13/0016 20130101;
G06K 19/06187 20130101 |
International
Class: |
G11C 13/00 20060101
G11C013/00; B01J 20/26 20060101 B01J020/26; C08L 101/00 20060101
C08L101/00; G06K 19/06 20060101 G06K019/06 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 29, 2016 |
AT |
A 50393/2016 |
Claims
1. A method for reading out stored information of a molecularly
imprinted polymer that comprises a defined sequence of different
functional groups with which a meaning of a defined value of a
numerical code is associated, comprising: bringing the molecularly
imprinted polymer into contact with a pool of analyte templates,
wherein the analyte templates have different side functionalities
that are complementary to the functional groups of the molecularly
imprinted polymer, and wherein the analyte templates differ from
one another with respect to an order of side functionalities
thereof, so that the analyte template that has a sequence of side
functionalities that is complementary to the functional groups
binds specifically to a sequence of different functional groups of
the numerical code of the molecularly imprinted polymer.
2. The method according to claim 1, wherein the pool of analyte
templates contains the template complementary to the sequence of
the functional groups as well as isomers, enantiomers, and/or
variants of the complementary template.
3. The method according to claim 1, wherein the analyte template of
the pool and monomers of the molecularly imprinted polymer that
comprise the functional groups are isotopically labelled.
4. The method according to claim 3, wherein the information is read
out by spatially-resolving dipolar solid state NMR.
5. A method for reading out stored information of a molecularly
imprinted polymer that is selectively provided with a defined
sequence of different functional groups reflecting a stored value
of a numerical code, wherein an anti-idiotypic method is used to
read out the stored value, the method comprising: producing a pool
of molecules that are template components having different side
functionalities, wherein each template component has one side
functionality that is complementary to one of the functional groups
of the defined sequence; bringing the pool into contact with the
molecularly imprinted polymer, wherein an imprint of the
molecularly imprinted polymer that comprises the stored value of
the numerical code acts as a reaction chamber, so that the template
components bind to the different functional groups of the imprint
according to respective side functionalities thereof, such that a
replica of the template used to produce the stored value of the
numerical code is created in the imprint; and reading out the
stored value by characterization of replicas by means of an
analytical method.
6. The method according to claim 5, wherein the template components
of the pool and monomers of the molecularly imprinted polymer that
comprise the functional groups are isotopically labelled.
7. The method according to claim 6, wherein the information is read
out by spatially-resolving dipolar solid state NMR.
8. A method for reading out the stored information of a molecularly
imprinted polymer that comprises a defined sequence of functional
groups with which the meaning of a defined value of a numerical
code is associated, comprising: bringing said molecularly imprinted
polymer into contact with a pool of template components or analyte
templates, which analyte templates are a sequence of said template
components; wherein said analyte templates differ from one another
with respect to an order of side functionalities thereof, wherein
said template components have different side functionalities that
are complementary to said functional groups of said molecularly
imprinted polymer, and wherein said template components or analyte
templates that have a sequence of side functionalities that is
complementary to said sequence of functional groups of the
numerical code of the molecularly imprinted polymer bind
specifically to said functional groups of said polymer.
9. The method according to claim 8, wherein said template
components or said analyte templates that are sequences of said
template components and monomers of the molecularly imprinted
polymer that comprise said functional groups are isotopically
labelled.
10. The method according to claim 9, wherein said stored
information is read out by spatially-resolving dipolar solid state
NMR.
11. The method according to claim 8, wherein said code is a binary
code, and two different template components and thus only two
different side functionalities are used.
12. A method for reading out the stored information of a
molecularly imprinted polymer that comprises a defined sequence of
functional groups with which the meaning of a defined value of a
numerical code is associated, by the use of template components,
wherein multiple template components are used that differ from each
other with respect to their side functionalities, said side
functionalities are complementary to said functional groups of said
molecularly imprinted polymer, the method comprising: bringing said
molecularly imprinted polymer into contact with a pool of template
components or multiple different sequences of said template
components, which sequences of said template components differ from
one another with respect to an order of side functionalities
thereof, wherein said template components bind specifically to said
defined sequence of functional groups of said polymer.
13. The method according to claim 12, wherein said template
components or said sequences of said template components and
monomers of the molecularly imprinted polymer that comprise the
functional groups are isotopically labelled.
14. The method according to claim 13, wherein said information is
read out by spatially-resolving dipolar solid state NMR.
15. The method according to claim 12, wherein said numerical code
is a binary code, and two different template components that differ
from each other with respect to their side functionalities are
used.
16. A method for retrieving information, comprising: forming a
molecularly imprinted polymer comprising a defined sequence of
functional groups, the functional groups being adapted to bind with
respective side functionalities of template components of
templates, the side functionalities of the template components
adapted to bind to a defined sequence of functional groups of the
molecularly imprinted polymer; and contacting the molecularly
imprinted polymer with a pool of templates having template
components, the templates differing from one another with respect
to an order of template components thereof, the functional groups
binding with the respective side functionalities, the template
having the template components ordered so that the side
functionalities are complementary to the functional groups of the
molecularly imprinted polymer binding with the molecularly
imprinted polymer, an order of the template components of the
template binding with the molecularly imprinted polymer defining a
stored value.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S. patent
application Ser. No. 16/096,008, filed Oct. 24, 2018, entitled
"STORAGE OF CODES IN MOLECULARLY IMPRINTED POLYMERS", which is a
national phase application of PCT Application No.
PCT/AT2017/060111, filed Apr. 27, 2017, which claims priority to
AT20160050393, dated Apr. 29, 2016, all of which are incorporated
by reference in their entirety.
BACKGROUND
[0002] 1) Field of the Invention
[0003] The invention relates to the storage of numerical codes,
more particularly binary codes, in a polymer structure based on a
molecularly imprinted polymer that is provided with complementary
imprints of a sequence of at least two different chemical
functionalities of a template.
[0004] 2) Description of Related Art
[0005] The ability to store digital information on conventional
hard drives and similar data carriers will reach its limit in the
near future, because the storage densities of such carriers cannot
be extended as required. Exponential growth in data volumes
requires the development of additional, alternative storage methods
or materials. Binary-encoded macromolecules represent here an
opportunity for long-term preservation of digital data.
[0006] The model for data storage in polymers can be found in
natural DNA. Sequences of nucleobases, linked to a phosphate- and
sugar molecular-based polymer backbone, carry a large volume of
information that can be translated into protein molecules. Such DNA
sequences may also be produced synthetically. Specific sequencing
of the nucleobases used makes it possible to represent and store
digitalized data such as text, images, and audio in binary code
("Towards practical, high-capacity, low-maintenance information
storage in synthesized DNA." Nature, 494: 77-80, 2013; "Robust
Chemical Preservation of Digital Information on DNA in Silica with
Error-Correcting Codes." Angew. Chem. Int. Ed., 54: 2552-2555,
2015).
[0007] Non-natural polymers such as poly(alkoxyamine amide)s can
also be used, however, to store digital information ("Design and
synthesis of digitally encoded polymers that can be decoded and
erased." Nat. Commun. 6: 7237, 2015).
[0008] In the face of DNA, and with non-natural polymers, the data
or codes are thus synthesized directly, wherein the synthesized
polymer itself acts as the data storage.
[0009] Also known from the prior art is the principle of molecular
imprinting of polymers (molecularly imprinted molecules (MIPs)).
Molecular imprinting is a technique developed, inter alia, by the
Mosbach group; see "Drug assay using antibody mimics made by
molecular imprinting." Nature 361: 645-647, 1993; "Molecularly
Imprinted Polymers and Their Use in Biomimetic Sensors." Chem. Rev.
100 (7): 2495-2504, 2000; "Molecular Imprinting: Synthetic
Materials as Substitutes for Biological Antibodies and Receptors.
Chem. Mater." 20 (3): 859-868, 2008; "Synthesis of
substrate-selective polymers by hostguest polymerization."
Makromol. Chem. 182 (2): 687-692, 1981; "New Configurations and
Applications of Molecularly Imprinted Polymers" J. Chromatogr. A,
889: 15-24, 2000; Bruggemann O (2002) "Molecularly imprinted
materials--receptors more durable than nature can provide." Chapter
in Advances in Biochemical Engineering/Biotechnology, Special
Issue: Modern Advances in Chromatography, Springer, edited by Prof.
Dr. R. Freitag.
[0010] Biomedical uses of MIPs are described by Liu et al. in
"Preparation of protein imprinted materials by hierarchical
imprinting techniques and application in selective depletion of
albumin from human serum." Sci Rep., 4:5487.doi:10.1038/srep05487,
2014 Jun. 30; by Ciardelli et al. in "The relevance of the transfer
of molecular information between natural and synthetic materials in
the realization of biomedical devices with enhanced properties." J
Biomater Sci Polym Ed., 16(2):219-36, 2005; and by Shi. et al. in
"Template-imprinted nanostructured surfaces for protein
recognition. Nature, 398(6728):593-7, 1999 Apr. 15.
[0011] WO 1995021673 A1 and the publication "Generation of new
enzyme inhibitors using imprinted binding sites: the anti-idiotypic
approach, a step toward the next generation of molecular
imprinting". J. Am. Chem. Soc., 123(49): 12420-12421, 2001 disclose
the use of anti-idiotypic methods for MIPs.
[0012] In molecular imprinting, first a template molecule is
selected. In particular, biomolecules, for example, vitamins,
hormones, or proteins are used as the template molecule. The
template molecule has, depending on the nature thereof, a plurality
of functional groups to which complementary functional groups can
bind. Because the functional groups of the template molecule have a
specific arrangement relative to one another, the template molecule
binds specifically only to another molecule that has the
complementary arrangement of the complementary functional groups.
In nature, signal molecules bind to receptors according to this
principle. In molecular imprinting, a receptor for the template
molecule is produced artificially, by bringing different functional
monomers having different functional groups into contact with the
template molecule, so that the monomers bind to the respective
complementary functional group on the template molecule. Doing so
does not require knowing the arrangement of the functional groups
on the template molecule, which plays no role in the process of
molecular imprinting. Once bonded to the template molecule, the
monomers are cross-linked to one another, such that the monomers
are fixed in the positions and orientations thereof relative to one
another, in order to form a polymer. The template molecule is then
removed, so that a molecular imprint of the template molecule stays
behind in the polymer and can consequently be used as an artificial
receptor for the template molecule, in particular, a biomolecule.
The information content of the imprint or the MIP is limited to
whether or not a biomolecule binds specifically, i.e., is limited
to either a YES or a NO. Molecularly imprinted polymers (MIPs) can
thus be put to use for specific recognition of the template in
chromatographic, extractive, or sensory applications.
SUMMARY OF THE INVENTION
[0013] This technique has thus far not been used to store data, or
to store digital information or codes.
[0014] The invention solves the problem of providing an improved
method for storing values or digital data at the molecular
level.
[0015] The invention solves this problem in that: in a first step,
a template molecule or template having a defined sequence of
defined functional groups is produced, the sequence representing a
defined value of a numerical code, preferably, a digital code, or
containing digital data; and in a second step, the defined sequence
of the template molecule is transferred according to the method of
the molecular imprinting to a polymer by bringing the template into
contact with monomers that have complementary functional groups and
therefore align themselves according to the sequence of defined
functional groups on the template, the monomers being successively
fixed to one another by polymerization so that the functional
groups henceforth carry the digital data. For improved clarity of
reading, the functional groups of the template in succession are
called side functionalities, in order to distinguish the
terminology therefor from that for the functional groups of the
monomers.
[0016] The data carriers according to the invention are thus
molecularly imprinted polymers (MIPs) that contain a defined
sequence of monomers or monomer units having defined functional
groups, wherein preferably the functional groups of one monomer
unit code for 0 and the functional groups of another monomer unit
code for 1. The information content thus lies in the sequence or
order of the functional groups on the MIP, and thus represents a
numerical code. The radix (number of different functional groups)
of the numerical code is preferably two, such that the code is a
binary code. If the template is produced with a defined sequence of
more than two different functional groups, such that monomers or
monomer units having more than two different functional groups bind
to the template, it is also possible to store a numerical code that
has a higher radix than 2 in the MIP according to the
invention.
[0017] It is advantageous that selecting the number of monomers and
preferably selecting a suitable cross-linker makes it possible to
produce molecularly imprinted polymer data carriers that have
different properties or shapes, which would not be possible when a
molecular data carrier is produced directly, e.g., as DNA.
[0018] Within the scope of the invention, therefore, templates
having defined sequences of template molecules, which are available
with different side functionalities, are produced first, these
sequences being carriers for the desired numerical code, more
particularly, binary code.
[0019] The template thus contains a sequence of at least two
template components each having different chemical side
functionalities, wherein these two different side functionalities
correspond to the binary numbers 0 and 1. It may then occur that a
template is composed of a sequence of only one template component,
if, for example, the code consists solely of the binary number 0,
or solely of the binary number 1. In a preferred embodiment, the
template is composed of a sequence of two template components each
having different chemical side functionalities, wherein the
sequence contains at least 3, 4, 5, 6, 10, 15, 20, or more
components.
[0020] Examples of possible template components include chemical
molecules that differ in the functional side chains thereof, in
particular, in the side functionalities thereof. Especially
suitable are those molecules that bear a carboxyl group or primary
amino group as a side functionality, preferably as terminal groups.
Other functional groups are also suitable as side functionalities,
however, provided said functional groups are able to form a
connection with a complementary group. As template components, it
is also possible to use: nucleotides; nucleotide derivatives such
as, for example, peptide nucleic acids; basic or acid vinyl
monomers; oligomerizable anionic or cationic monomer units and
other chemically linkable structural units each having additional
side functionalities, such as, for example, omega-hydroxycarboxylic
acids with an additional carboxy or amino function, or omega-amino
acids with an additional carboxy or amino function.
[0021] Examples of especially suitable templates include peptides
and proteins that are composed of two different amino acids as
template components. Preferably, one template component is an acid
amino acid, and the other template component is a basic amino acid.
The different enantiomers of these molecules may then also be
used.
[0022] Peptide nucleic acid (PNA) structures composed of a sequence
of two different nucleobase components are also suitable as
templates. With such peptide nucleic acids, the sugar phosphate
backbone is replaced, for example, with a pseudopeptide.
[0023] With the help of the template or at least one template
having a defined sequence of side functionalities, a polymer is
imprinted according to the invention.
[0024] The method according to the invention for producing a
molecularly imprinted polymer is performed by imprinting the
polymer of the molecularly imprinted polymer by polymerizing the
polymer in the presence of at least one template, wherein the
template is composed of a defined, selected sequence of structural
components, wherein the structural components are selected from at
least two types of structural components that differ from one
another at least with respect to the side functionalities thereof,
wherein templates having any sequence of the structural components
thereof--i.e., according to any value of the numerical code--can be
produced, wherein at the side functionalities of the template,
monomers are bonded with the functional groups thereof that are
complementary to the side functionalities, wherein the monomers
differ from one another with respect to the functional groups
thereof, and wherein the monomers are bonded when the
polymerization takes place in the polymer structure of the polymer,
and the template is subsequently released with the side
functionalities thereof from the monomers, so that the molecularly
imprinted polymer comprises a defined value of the numerical code,
more particularly, the binary code, formed of the functional groups
of successive monomers, corresponding to the selected sequence of
the structural components of the template that was used to produce
the molecularly imprinted polymer.
[0025] Preferably, the monomers--more particularly, the functional
groups thereof--are isotopically labelled.
[0026] The invention comprises molecularly imprinted polymers
(MIPs) containing a defined value of a numerical code, more
particularly, a binary code, that are produced according to the
method according to the invention.
[0027] A preferred embodiment of the invention comprises a
molecularly imprinted polymer (MIP) containing a binary code,
wherein the molecularly imprinted polymer (MIP) contains a defined
sequence of monomers, wherein the functional group of one monomer
codes for the binary number 0 and the functional group of another
monomer codes for the binary number 1.
[0028] Another embodiment of the invention comprises a molecularly
imprinted polymer (MIP) containing a numerical code, more
particularly, a binary code, wherein the monomers of the
molecularly imprinted polymer (MIP) differ from one another with
respect to the functional groups thereof.
[0029] Another preferred embodiment of the invention comprises a
molecularly imprinted polymer (MIP) containing a binary code,
wherein one monomer or monomer unit of the MIP has an acidic group,
e.g., a carboxyl group, and the other monomer or other monomer unit
of the MIP comprises a basic group, e.g., an amino group.
[0030] Another preferred embodiment of the invention comprises a
molecularly imprinted polymer (MIP) containing a binary code,
wherein one monomer of the MIP is methacrylic acid and the other
monomer is 2-aminoethyl methacrylate.
[0031] Another embodiment of the invention comprises a molecularly
imprinted polymer (MIP) containing a numerical code, more
particularly, a binary code, wherein the sequence of the monomers
has at least a length of three monomers, wherein the monomers may
be identical or different.
[0032] A sequence or stored numerical value may preferably have a
length of at least 3, 5, 8, 10, 15, 20, 25, 30, 50 monomers.
[0033] Another preferred embodiment of the invention comprises a
molecularly imprinted polymer (MIP) containing a binary code,
wherein the sequence has a length of at least three monomers,
wherein at least one monomer of the sequence bears a carboxyl group
and at least one monomer of the sequence bears an amino
function.
[0034] Preferably, the molecularly imprinted polymer is produced
according to the following steps: [0035] a. producing the template,
the template being produced as a freely-definable sequence of
template components having different chemical side functionalities,
wherein one template side functionality can be recognized as
logical 1 and one template side functionality can be recognized as
logical 0; [0036] b. adding the monomers, which have complementary
functional groups to the side functionalities of the template;
[0037] c. self-organization by the monomers at the side
functionalities of the template components via the complementary
functional groups thereof; [0038] d. fixing the complementary
binary code by polymerizing the monomers in order to produce the
polymer; and [0039] e. removing the template from the polymer so
that the functional groups of the monomer units are exposed, such
that the polymer exists as a molecularly imprinted polymer, wherein
the sequence of the functional groups forms the defined value of
the binary code.
[0040] The complementary monomers, i.e., the monomers added in step
b) may be selected, for example, from anionic and cationic
monomers. Examples of anionic monomers include monomers having
electron-withdrawing substituents, such as nitrile, carboxyl,
phenyl, and vinyl groups, such as acrylic acid, methacrylic acid,
crotonic acid, itaconic acid, fumaric acid, maleic acid, monomethyl
itaconate, monomethyl fumarate, monobutyl fumarate, maleic
anhydride, acrylamido glycolic acid, styrenesulfonic acid,
vinylsulfonic acid, vinylphosphonic acid,
2-acrylamido-2-methylpropane phosphonic acid,
2-acrylamido-2-methyl-1-propanesulfonic acid, and derivatives of
the anionic monomers mentioned in this paragraph.
[0041] Examples of cationic monomers include--but are not limited
to--cationic ethylenically unsaturated monomers such as
diallyldialkylammonium halides such as diallyl dimethyl ammonium
chloride, the (meth)acrylates of dialkylaminoalkyl compounds such
as dimethylaminoethyl (meth) acrylate, diethylaminoethyl
(meth)acrylate, dimethylaminopropyl (meth)acrylate,
2-hydroxydimethylaminopropyl (meth)acrylate, aminoethyl
(meth)acrylate, and salts and quaternary compounds thereof,
N,N-dialkylaminoalkyl (meth)acrylamide such as
N,N-dimethylaminoethyl acrylamide and salts and quaternary
compounds thereof, and derivatives of the cationic monomers
mentioned in this paragraph.
[0042] Suitable complementary monomers thus contain complementary
functional groups.
[0043] The molecularly imprinted polymer is preferably
biodegradable.
[0044] This invention also comprises methods for reading out the
code of a molecularly imprinted polymer according to the
invention.
[0045] A first method according to the invention for reading out
the stored information of a molecularly imprinted polymer that has
a defined sequence of different functional groups reflecting a
defined value of a numerical code, more particularly, a binary
code, is performed by bringing the molecularly imprinted polymer
into contact with a pool of analyte templates, wherein the analyte
templates have different side functionalities that are
complementary to the functional groups of the molecularly imprinted
polymer, wherein the analyte templates differ from one another with
respect to the order of side functionalities thereof, so that only
that analyte template that has the sequence of side functionalities
that is complementary to the functional groups binds specifically
to a sequence of different functional groups of the numerical code
of the molecularly imprinted polymer.
[0046] Thus, that analyte template that corresponds to the template
that was used to produce the molecularly imprinted polymer analyte
template binds specifically to the molecular imprint of the
molecularly imprinted polymer according to the invention.
Preferably, the analyte templates of the pool have been
isotopically labelled.
[0047] A second method according to the invention for reading out
the stored information of a molecularly imprinted polymer that is
selectively provided with a defined sequence of different
functional groups reflecting a defined value of a numerical code,
more particularly, a binary code, is performed by using an
anti-idiotypic method to read out the stored information, the
method comprising the steps of: [0048] a. producing a pool of
molecules that are template components having different side
functionalities, wherein each type of template component has one
side functionality that is complementary to one of the functional
groups of the defined sequence; [0049] b. bringing the pool of
template components into contact with the molecularly imprinted
polymer, wherein the imprint of the molecularly imprinted polymer
that has the stored value of the numerical code acts as a reaction
chamber, so that template components bind to the different
functional groups of the imprint according to the respective side
functionalities thereof, such that a replica of the template that
may have been used or was used to produce the stored value of the
numerical code is created in the imprint; and [0050] c. reading out
the stored value by characterization of replicas by means of an
analytical method.
[0051] The second method according to the invention for reading out
the value of the code of the MIP thus differs from the first in
that only the template components are used, instead of templates
composed of bonded template components. This is advantageous in
that it is not necessary to produce all of the relevant variants of
analyte templates and bring the same into contact with the
molecularly imprinted polymer, but rather only the template
components from which the templates or analyte templates were
formed. In the case of a binary code, thus, only two different
template components are required in the second readout method
according to the invention.
[0052] Preferably, the template components of the pool have been
isotopically labelled.
BRIEF DESCRIPTION OF THE DRAWINGS
[0053] The drawings provide a more detailed illustration, by way of
example, of the method according to the invention on the basis of
several embodiment variants. The drawings show:
[0054] FIG. 1. an exemplary template molecule having a binary code
based on two different template components (lysine and glutamic
acid);
[0055] FIG. 2. an exemplary template molecule in electrostatic
interaction with corresponding, complementary functional
monomers;
[0056] FIG. 3. the molecular imprint of the exemplary template
molecule in the polymer, wherein the template molecule is still
embedded;
[0057] FIG. 4. the molecular imprint of the exemplary template
molecule in the polymer, after the template molecule has been
removed;
[0058] FIG. 5. the specific recognition of the binary code stored
in the molecular imprint of the polymer, through the template
molecule; and
[0059] FIG. 6. the reading out of the stored binary code through
spatially-resolved solid state NMR in the molecular imprint.
Selective isotopic labelling of the molecular imprint or the
monomer units and template molecules enables magnetic dipolar
interactions.
[0060] In the chemical structural formula, "PG" stands for
"protecting group."
DETAILED DESCRIPTION
[0061] The octapeptide lysine-lysine-lysine-lysine-lysine-glutamic
acid-glutamic acid-lysine is used as a template 1, by way of
example (see FIG. 1). With lysine and glutamic acid, the template 1
has two different template components that differ from one another
with respect to the side functionalities 2, 3 thereof. In the
example of FIG. 1, the amino function, as the side functionality 2
of the lysine, acts as a binary "1" while the carboxyl function, as
the side functionality 3 of the glutamic acid, acts as a binary
"0." The following sequence of side functionalities 2, 3--read from
left to right--thus arises for the example template 1 depicted in
FIG. 1: Amino-Amino-Amino-Amino-Amino-Carboxy-Carboxy-Amino, thus
11111001 as the value of the binary code 4.
[0062] The addition of at least two different monomers 5 having
functional groups 6, 7 that are complementary to the side
functionalities 2, 3 of the template 1 is followed then by a wait
for the self-organization of the template 1 and monomers 5 via the
functionalities thereof, so that the monomers 5 bind to the side
functionalities 2, 3 according to the functional groups 6, 7
thereof, as is illustrated in FIG. 2.
[0063] Suitable monomers 5 thus contain complementary functional
groups. Thus, as illustrated in the example, the first monomer
5--methacrylic acid, with the functional group 6 thereof--is
complementary to the side functionality 2 in the form of the amino
function of the template component lysine. The second monomer 5, in
the form of 2-aminoethyl methacrylate with the functional group 7
thereof in the form of an amino-functionalized side chain, is
complementary to the side functionality 3 in the form of the
carboxy function of the template component glutamic acid. Thus, the
functionally complementary monomers 5 organize themselves with the
template components through the complementary functional groups
thereof. In the example of FIG. 2, the acid function of the
methacrylic acid organizes itself with the basic function of the
lysine, and the basic function of the 2-aminoethyl methacrylate
organizes itself with the acidic function of the glutamic acid of
the octapeptide that forms the template 1. Electrostatic
interactions thus form stable compounds.
[0064] After a cross-linking monomer has been added, the monomers 5
can be polymerized to thereby fix and store the complementary
template structure and thus the binary code 4. Examples of suitable
monomeric cross-linkers include ethylene glycol dimethacrylate,
butylene glycol dimethacrylate (or butane-1,4-diol dimethacrylate),
and hexamethylene dimethacrylate (or hexane-1,6-diol
dimethacrylate).
[0065] In FIG. 3, the monomers 5 are depicted in the cross-linked
state thereof, i.e., the monomers are components of a polymer 8 or
are bonded to a polymer 8. As is represented, the template 1 is
still bonded to the polymer 8 after the polymerization.
[0066] As depicted in FIG. 4, the template 1 is removed from the
polymer 8. After the template 1 has been removed with the original
binary code 4 thereof, then, the functionalities of the previous
monomers 5 remain behind in the resulting molecular imprints of the
polymer 8 in an immobilized configuration, such as the template 1
was set forth. The polymer 8, after the template 1 has been
removed, thus exists as the molecularly imprinted polymer 9. The
molecularly imprinted polymer 9 has, in an exposed state, the
functional groups 6, 7 of the cross-linked monomers 5, which form
the binary code 4 of the molecularly imprinted polymer 9, i.e.,
according to the example, the code sequence or the stored value
11111001.
[0067] As illustrated in FIG. 5, the thus-stored value of the
binary code 4 is read out from the molecular imprints of the MIPs 9
according to a first variant according to the invention by stirring
a mixture of analyte templates 10 also containing an analyte
template 10 that corresponds to the original template 1 with a
suspension of MIP particles, and measuring the residual content of
the analyte templates 10 in the supernatant of the MIP particles
after an adsorption phase. The content of the analyte template 10
that corresponds to the original template 1 is diminished in
comparison to the other analyte template 10 because that analyte
template 10 binds specifically as the sole molecule in the
molecular imprints. This makes it possible to determine which value
of the binary code 4 is present in the molecular imprint of the MIP
9. As shown, the analyte template 10 that has the sequence of side
functionalities 2, 3 that is complementary to the sequence of the
functional groups 6, 7 of the MIP 9 binds specifically to the
imprint, i.e., the sequence 22222332 binds specifically to the
sequence 66666776, i.e., according to the example, the side
functionality sequence
Amino-Amino-Amino-Amino-Amino-Carboxy-Carboxy-Amino of the template
component sequence Lysine-Lysine-Lysine-Lysine-Lysine-Glutamic
acid-Glutamic acid-Lysine binds to the functional group sequence
Carboxy-Carboxy-Carboxy-Carboxy-Carboxy-Amino-Amino-Carboxy of the
monomers 5.
[0068] According to a second readout method according to the
invention, the binary code 4 may also be read out by adding
solutions of chemical structural components of the original
template molecules according to a type of anti-idiotypic method and
then replicating these template molecules in the molecular
imprints, it being possible to determine the code thereof after
elution and analytical characterization. This second readout method
according to the invention is thus performed by producing a pool of
molecules that contain at least the original template components of
the template 1 that was used to produce the MIP 9. This pool is
brought into contact with the MIP 9, wherein the molecular imprint,
i.e., the binary code 4, of the MIP 9 acts as a reaction chamber.
The complementary template components of the pool bind to the
molecular imprint, thereby producing replicas of the original
templates 1. These replicas may be characterized by means of
analytical methods, for example, by means of chromatographic
methods, and thus the stored code 4 can be read out. The molecular
imprint in the MIP 9 may act, on the one hand, as a copy room for
replicating the original template 1, while the molecular imprint
may also be used, on the other hand, to produce different variants
or derivatives of the original template 1, depending on the choice
of chemical components, with an unaltered sequence of the side
group functionalities, i.e., of the binary code 4. In other words,
the code in the MIP 9 can be used to produce duplicates or
derivatives of the template 1, which can be used in turn as data or
information carriers, or can be used to produce other MIPs 9. The
MIPs 9 according to the invention can thus be copied or
replicated.
[0069] FIG. 6 illustrates a method for reading out the stored
binary code directly at the MIP 9 by spatially-resolving solid
state NMR. NMR stands for nuclear magnetic resonance. It is to be
provided according to the invention that the monomers 5 and the
template component binding thereto have a selective isotopic
labelling, which is achieved, for example, by selecting the
nitrogen atoms of the amino functions and the carbon atoms of the
carboxy functions of both the side functionalities 2, 3 of the
template components and the functional groups 6, 7 of the monomers
5 in the form of .sup.15N and .sup.13C isotopes, respectively. With
spatially-resolving, multi-nuclear, multi-dimensional solid state
NMR, the binary code 4 is read out according to the invention by
measuring a dipolar recoupling by means of rotational echo double
resonance (REDOR) or radio frequency-driven recoupling (RFDR)
spectroscopy on the basis of the aforementioned isotopic labelling,
and thus being able to determine the structure and orientation of
the template 1, or an identical analyte template 10, in the MIP 9
and thus the order of the functional groups 6, 7 in the MIP 9 and
therewith the value of the binary code 4.
[0070] The isotopically labelled template components in the MIP 9
may, on the basis of the first readout method according to the
invention, be bonded by bringing a pool of different isotopically
labelled analyte templates 10 differing from one another in the
order of the isotopically labelled template components thereof in
contact with the MIP 9, so that only that isotopically labelled
analyte template 10 that has the value of the binary code 4 of the
original template 1 binds to the imprint of the MIP 9, as is
illustrated in FIG. 6.
[0071] The second readout method according to the invention--which
follows a type of anti-idiotypic method--may preferably be carried
out with isotopically labelled template components. The
isotopically labelled template components bind with the respective
side functionalities 2, 3 thereof to the complementary functional
groups 6, 7 of the imprint, i.e., according to the order of the
binary code 4, such that the isotopically labelled template
components together form a duplicate or derivative of the original
template 1, which exists according to the analyte template 10 of
FIG. 6 in the isotopically labelled imprint of the MIP 9.
[0072] Because the measurable interaction between the isotopes of
the bonded analyte template 10 and the isotopes of the monomers 5
differ according to the order of the arrangements thereof, the
value of the binary code 4 can be determined directly at the MIP
9.
[0073] The molecularly imprinted polymers 9 described herein are
produced in the presence of the template 1, preferably via a
surface, precipitation, suspension, emulsion, or mass
polymerization in a batch or semi-batch process, and put to use in
different forms, preferably in the form of spherical particles,
or--especially preferably--in the form of polymer coatings.
[0074] The spherical particles or polymer coatings may be used, for
example, to encode for products of every kind. Due to the size down
to the nanometer range, the MIPs 9 are invisible to the consumer
when applied to long-lasting products, so that the origin thereof
can be unambiguously determined even after a long period of time
has passed. The MIPs 9 can thus contain, for example, detailed
information on the actual origin of the original products, so that
the products can be distinguished from counterfeits. Plastic
matrices may be provided directly with the described molecular
imprints and thus be encoded or generally put to use as data
carriers. For example, specific manufacturer or customer data, or
simply the date of production, may be left as a numerical value or
in binary form in the imprint.
[0075] It is also possible to produce multi-MIPs 9, wherein a
plurality of different templates 1 are used, in order to imprint,
in parallel, different numerical codes, more particularly, binary
codes 4 having different information into molecular imprints. One
MIP 9 can thus comprise a plurality of different molecular
imprints, which may differ from one another with respect to the
code sequence and/or code length thereof.
[0076] Thus, another embodiment comprises MIPs 9 that contain at
least two different values of a numerical code, more particularly,
a binary code 4.
[0077] In one embodiment of the invention, the MIPs 9 are used to
recognize and/or code for foodstuffs, consumer goods, industrial
goods, and components or ingredients thereof.
Example 1
[0078] To produce a molecularly imprinted polymer 9 according to
the invention as an example, the tripeptide glutamic
acid-lysin-lysine (EKK) was used as the template 1. The value of
the binary code 4 present in the amino acid sequence corresponds
thus to 100. The formulation of this template polymer is set forth
in table 1.
TABLE-US-00001 TABLE 1 Composition of the molecularly imprinted
polymer (MIP1) with use of the template EKK, with molar mass,
calculated and actually- measured mass of the substances, and the
equivalents thereof Molar Substance mass g/mol Estimated Actual
Equivalent Template EKK 625.31 15 mg 15.67 mg 1 Methacrylamide
86.04 33.00 mg 33.54 mg 15.6 Methacrylic acid 85.05 32.64 mg 33.65
mg 15.8 Ethylene glycol 198.22 237.75 mg 237.38 mg 47.8
dimethacrylate Azobisisobutyr- 164.21 1.17 mg 1.42 mg 0.35 onitrile
Acetonitrile 41.05 3.75 mL 3.75 mL -- Dimethylsulf- 78.13 -- 0.2 mL
-- oxide
[0079] With the exception of the initiator azobisisobutyronitrile,
all of the components were dissolved in a mixture of acetonitrile
and dimethyl sulfoxide. The solution was stirred for 4 hours in
order to make it possible to create electrostatic interactions such
as hydrogen bonds and--in addition, after proton transfer--ionic
bonds between the template 1 and the functional monomers 5
methacrylamide and methacrylic acid. The initiator
azobisisobutyronitrile is then added thereto, and the solution was
sprayed for 5 minutes with gaseous nitrogen. Then, in a
refrigerator at 6.degree. C., the solution was placed in a UV
reactor and subjected to 24 hours of UV radiation. The suspension
formed was subsequently stirred for 24 hours with 6 mL of a
methanol-acetic acid mixture (9:1, v:v), in order to purify the
polymer 8 and, in particular, to remove the template molecules. The
resulting molecularly imprinted polymer 9 was then filtered and
washed twice with a methanol-acetic acid mixture and four times
with acetonitrile. The molecularly imprinted polymer 9 was
subjected to 5 more minutes of suction as a first round of drying.
Further drying steps included spraying the solid with gaseous
nitrogen for 5 minutes, and depositing in a drying oven at
40.degree. C. for a period of 24 hours. The yield of the
white-colored, powdery molecularly imprinted polymer 9 was 219.66
mg.
[0080] The template 1 (the tripeptide EKK) as analyte and other
comparison analytes/analyte templates 10 (the tripeptides KEK, EKE,
EEK, EEE) were each dissolved in 0.1 mL of dimethyl sulfoxide and 8
mL of acetonitrile, and the powdery MIP 9 was suspended therein.
Table 2 lists the exact details of these affinity assays. These
suspensions were each stirred for 18 hours at room temperature. 2
mL was then removed from each of these suspensions and centrifuged
at a rotational speed of 10,000 RPM. The resulting supernatants
were diluted with 8 mL of acetonitrile and the solutions were then
subjected to spectroscopic measurement at a wavelength of 300
nm.
TABLE-US-00002 TABLE 2 Affinity assays with the molecularly
imprinted polymer MIP1 with different tripeptides, the absolute
masses used thereof, the masses of the molecularly imprinted
polymer MIP1 used, and the measured concentrations of the
tripeptides in the supernatant after reaching equilibrium.
Concentration in supernatant based on Analyte (peptide Mass of Mass
of measured absorption sequence) analyte/mg MIP/mg C.sub.calc,
mg/mL KEK 2.11 10.43 0.073 EKK* 2.12 10.39 0.043 EKE 2.08 10.20
0.069 EEK 2.10 9.94 0.072 EEE 2.16 10.31 0.117 (E = glutamic acid,
K = lysine) *corresponds to the original template molecule
[0081] This example showed that the MIP 1 has a particular affinity
to the original template EKK (line marked with *), with an
especially high adsorption due to specific molecular imprints, or
with an especially low residual content in the supernatant of only
0.043 mg/mL, in comparison to the four other tripeptides KEK (0.073
mg/mL), EKE (0.069 mg/mL), EEK (0.072 mg/mL), and EEE (0.117
mg/mL). In this manner, it was possible to read back, from a key
set of five tripeptide molecules (KEK, EKK, EKE, EEK, and EEE), the
matching key (EKK) due to the stored information, i.e., the
sequence EKK or the binary code 100.
* * * * *