U.S. patent application number 11/212454 was filed with the patent office on 2007-03-01 for method and device for preparing a biological sample for biological analyses.
Invention is credited to Goran Palmers, Karl Skold, Marcus Svensson.
Application Number | 20070048877 11/212454 |
Document ID | / |
Family ID | 37771859 |
Filed Date | 2007-03-01 |
United States Patent
Application |
20070048877 |
Kind Code |
A1 |
Skold; Karl ; et
al. |
March 1, 2007 |
Method and device for preparing a biological sample for biological
analyses
Abstract
The method and devices are for preparing a biological sample for
biological analyses. The biological sample has at least one amino
acid sequence from an organism. The amino acid sequence is a
subject of degradation. The biological sample is physically
prepared to enable altering a secondary structure of the amino acid
sequence to a deactivated secondary structure, and altering the
secondary structure of the amino acid sequence to a deactivated
secondary structure by heating the biological sample.
Inventors: |
Skold; Karl; (Uppsala,
SE) ; Svensson; Marcus; (Uppsala, SE) ;
Palmers; Goran; (Askim, SE) |
Correspondence
Address: |
FASTH LAW OFFICES (ROLF FASTH)
26 PINECREST PLAZA, SUITE 2
SOUTHERN PINES
NC
28387-4301
US
|
Family ID: |
37771859 |
Appl. No.: |
11/212454 |
Filed: |
August 26, 2005 |
Current U.S.
Class: |
436/174 |
Current CPC
Class: |
G01N 1/44 20130101; Y10T
436/25 20150115; C07K 1/1136 20130101 |
Class at
Publication: |
436/174 |
International
Class: |
G01N 1/00 20060101
G01N001/00 |
Claims
1. A method for preparing a biological sample for biological
analyses, the biological sample having an one amino acid sequence
from an organism, the amino acid sequence being a subject of
degradation, the method comprising: physically preparing the
biological sample to enable altering a secondary structure of the
amino acid sequence to a deactivated secondary structure, and
altering the secondary structure of the amino acid sequence to a
deactivated secondary structure by heating the biological
sample.
2. The method according to claim 1, wherein the method further
comprises step of physically preparing comprises shaping the
biological sample to enable altering a secondary structure of the
amino acid sequence to a deactivated secondary structure.
3. The method according to claim 2, wherein the method further
comprises step of shaping the biological sample comprises creating
a volume comprising the biological sample.
4. The method according to claim 3, wherein the method further
comprises step of creating the volume by filling the volume with a
filler presenting a similar dielectric constant as the biological
sample.
5. The method according to claim 4, wherein the method further
comprises step of the heating is microwave heating.
6. The method according to claim 3, wherein the method further
comprises step of creating the volume by designing the shape to
present a large heat transfer surface in relation to the volume of
the shape.
7. The method according to claim 6, wherein the method further
comprises step of creating the volume by shaping the biological
sample to a slice.
8. The method according to claim 6, wherein the method further
comprises step of the creating the volume by providing a thin and
oblong shape to the biological sample.
9. The method according to claim 2, wherein the method further
comprises step of heating by contact heat conduction.
10. The method according to claim 9, wherein the contact heat
conduction is done by at least of one gas, condensing gas, a fluid
and at least one warm plate transferring heat into the biological
sample.
11. The method according to claim 2, wherein the method further
comprises step of heating by using radiation.
12. The method according to claim 7, wherein the method further
comprises step of creating the volume by cutting the biological
sample.
13. The method according to claim 7, wherein the method further
comprises step of creating the volume by pressing the biological
sample.
14. The method according to claim 2, wherein prior to creating the
volume, freezing the biological sample.
15. The method according to claim 14, wherein after freezing the
biological sample, warming the biological sample to a sub zero
temperature allowing cutting the sample.
16. The method according to claim 15, wherein after warming the
biological sample, cutting the biological sample.
17. The method according to claim 6, wherein the method further
comprises step of creating the volume by placing the biological
sample in a container, vacuuming the container, and sealing the
container.
18. The method according to claim 6, wherein the method further
comprises step of creating the volume by leading the biological
sample into a tube, presenting heating means for heat transfer into
the biological sample.
19. The method according to claim 2, wherein the heating exceeds a
temperature similar to the temperature at which the secondary
structure of the amino acid sequence is altered to the deactivated
secondary structure.
20. A device for preparing a biological sample for biological
analyses, the biological sample has at least one amino acid
sequence from an organism, the one amino acid sequence is being a
subject of degradation, comprising: a power transfer location for
receiving the biological sample, and heating means for altering the
secondary structure of the amino acid sequence to a deactivated
secondary structure, the heating means being responsive for heating
the power transfer location.
21. Device according to claim 20, wherein the biological sample has
a container.
22. The device according to claim 20, wherein the power-transfer
location is constituted by a chamber.
23. The device according to claim 22, wherein the chamber presents
one of a circular cross section, a rectangular cross section, and a
triangular cross section.
24. The device according to claim 22, wherein the device has an
opening defined therein.
25. The device according to claim 24, wherein the device has a
second opening defined therein for allowing the biological sample
to exit the chamber.
26. The device according to claim 21, wherein an inner wall of the
chamber is essentially in contact with the container, when the
container is inserted into the chamber.
27. The device according to claim 21, wherein the container is one
of a needle and a tube.
28. The device according to claim 24, wherein the opening presents
a sealing, allowing insertion of the container into the
chamber.
29. The device according to claim 22, wherein the chamber has a
plurality of openable matching parts and a locking means for
maintaining the plurality of openable matching parts in a locked
state.
30. The device according to claim 22, wherein the device has a
steam generator that has an inlet defined therein and the chamber
has a steam inlet defined therein.
31. The device according to claim 20, wherein the power-transfer
location is constituted by at least one plate.
32. The device according to claim 20, wherein the heating means is
one of a radiation generator and a micro wave generator.
33. The device according to claim 20, wherein the power-transfer
location presents a large power transfer surface.
Description
TECHNICAL FIELD
[0001] According to a first aspect, the present invention relates
to a method for preparing a biological sample for biological
analyses. According to a second aspect, the present invention
relates to a device for preparing a biological sample for
biological analyses.
BACKGROUND OF INVENTION
[0002] The primary structure deals with the sequence of amino acids
and the secondary structure is defined by the phi and psi angles of
the backbone atoms of the amino acid residues, and the hydrogen
bonds between main chain atoms. In some cases, these dihedral
angles and patterns of hydrogen bonds are repeated throughout
subsequences of several consecutive residues, giving rise to
secondary structures, e.g. alpha helices and beta sheets.
[0003] A tertiary structure concerns how the secondary structure
units associate within a polypeptide chain providing a three
dimensional structure. A quaternary structure describes how at
least two polypeptide chains associate to form a native protein
structure, even though there are proteins that consist of a single
polypeptide chain.
[0004] Proteins and peptides have been widely investigated by
methods such as two dimensional gels and mass spectrometry.
Post-mortem activity of proteases and oxidative stress has been
shown to play an important role on peptide and protein
concentration in the brain, as well as for detecting
post-translational modifications. Previous studies have shown that
the peptide and protein content in brain tissue is affected by the
time-interval from death to the inactivation of proteases.
SUMMARY OF INVENTION
[0005] According to the first aspect, a method for preparing a
biological sample for biological analyses is disclosed. The
biological sample comprises at least one amino acid sequence from
an organism, and it is a subject of degradation. The method
comprises physically preparing the biological sample to enable
altering a secondary structure of the at least one amino acid
sequence to a deactivated secondary structure, and altering the
secondary structure of the at least one amino acid sequence to a
deactivated secondary structure by heating the biological sample.
Physically preparing implies, in an embodiment, that the biological
sample is given a shape that is preferred for facilitating an
effective heating in terms of the heating being uniform and fast.
This helps to shorten the time needed to reach a deactivated
secondary structure. By blocking biological processes driven by
proteins degradation of a sample is avoided. Since the time between
taking the biological sample and the performing a biological
analysis has a large impact on the level of degradation, even after
a short time, e.g. already after 1-3 min, it is important that
heating takes place immediately after taking the sample. By heating
the tissue protein functioning as proteases, their secondary and
tertiary structure and thereby their function is lost.
[0006] The method preserves the primary structure of proteins and
peptides by altering their original secondary, and their tertiary
structure. The heating of the tissue therefore has several
advantages; it enables the relatively low-abundant neuropeptides
and proteins to remain intact. In addition it minimizes degradation
of neuropeptides and proteins in a reproducible manner. This method
also makes it possible to compare the content and levels of
proteins and peptides from different samples. Also, the method may
be non fatal, i.e. the organism does not have to perish as a
consequence of using the method.
[0007] Non-limiting examples of biological samples constitute
biopsies from tissue such as muscles, skin, brain, liver, kidney,
or others or body fluids such as blood, urine, CSF, or others, or
cell extracts such as cell cultures.
[0008] In embodiments, heating the biological sample to a
temperature of the following ones is done: 50, 60, 70, 80, or 90
degrees Centigrade. In an embodiment, the heating reaches or
exceeds a temperature similar to the temperature at which the
secondary structure of an amino acid sequence is altered to the
deactivated secondary structure.
[0009] In an embodiment, the step of physically preparing comprises
shaping the biological sample to enable altering a secondary
structure of the at least one amino acid sequence to a deactivated
secondary structure, leading to effective heat transfer. In an
embodiment, shaping the biological sample comprises creating a
volume comprising the biological sample. Another advantage is that
when shaping the biological sample, the biological sample is given
a shape that is preferred for the heating, which should be quick
and uniform in order to stop the degradation in the whole sample as
soon as possible after taking the biological sample. It should be
pointed out that the volume may comprise only the biological
sample.
[0010] In an embodiment, creating the volume comprises filling the
volume with filler presenting similar electric and/or thermal
properties, such as heat capacity, as the biological sample. This
allows shaping, i.e. creating the volume, so that the heat is
transferred into the biological sample without discontinuities in
the contact between the filler and the biological sample, i.e. it
appears that the filler and the biological sample is constituted by
materials presenting similar electric properties. In an embodiment,
the filler is inert. This offers a basis for a uniform heating
process after which the whole biological sample is heated. In an
embodiment, creating the volume comprises filling the volume with
filler presenting similar light characteristics in terms of
refractive indices and absorption coefficients as the biological
sample. This allows shaping, i.e. creating the volume, so that the
heat is transferred into the biological sample without
discontinuities in the contact between the filler and the
biological sample, i.e. it appears that the filler and the
biological sample is constituted by materials presenting similar
light properties. This offers a basis for a uniform heating process
after which the whole biological sample is heated.
[0011] In an embodiment, the heating is microwave heating. In an
embodiment, creating the volume comprises designing the shape to
present a large heat transfer surface in relation to its volume.
This leads to an advantageous shape since a large surface allows
more heating effect to be received by the biological sample. Also,
this leads to that the longest distance in an unheated point in the
biological sample is short. This leads to an opportunity of
stopping, or at least delaying the degradation of the biological
sample even faster.
[0012] In an embodiment, creating the volume comprises shaping the
biological sample to a slice. In an embodiment, creating the volume
comprises providing a thin and oblong shape to the biological
sample. In an embodiment, heating is contact heat conduction. In an
embodiment, the contact heat conduction is done by at least of one
gas, condensing gas, a fluid and at least one warm plate
transferring heat into the biological sample. In an embodiment, the
heating is heating caused by radiation. In an embodiment, creating
the volume includes cutting the biological sample. In an
embodiment, creating the volume includes pressing the biological
sample. In an embodiment, prior to creating the volume, freezing
the biological sample is done for instance in order to make it
easier to cut a slice of the biological sample.
[0013] In an embodiment, after freezing the biological sample,
warming the biological sample to a sub zero temperature allowing
cutting the sample. In an embodiment, after warming the biological
sample, cutting the biological sample.
[0014] Having so far dealt with biological samples that present a
more solid characteristic, a few embodiments dealing with less
solid characteristics will now be presented.
[0015] In an embodiment, creating the volume comprises, in case the
biological sample presents a coagulating or a fluid characteristic,
placing the biological sample in a container, vacuuming the
container, and sealing the container. This offers the advantage of
being able to handle coagulating biological samples. In an
embodiment, creating the volume comprises, in case the biological
sample presents a fluid characteristic, leading the biological
sample into, or alternatively through, a tube, presenting heating
means for heat transfer into the biological sample. In embodiments,
the tube may be replaced by a flat passage, or a whirl canal.
[0016] According to a second aspect, a device for preparing a
biological sample for biological analyses is disclosed. The
biological sample comprises at least one amino acid sequence from
an organism. The at least one amino acid sequence is a subject of
degradation. The device comprises a power transfer location for
receiving the biological sample and heating means for altering the
secondary structure of the at least one amino acid sequence to a
deactivated secondary structure. The heating means is responsive
for heating the power transfer location. The power transfer
location receives power from the heating means and transfers power
to the biological sample and as a consequence the biological sample
is heated. Due to similarities with the first aspect, reference is
hereby made to the first aspect.
[0017] In an embodiment, the power transfer location is constituted
by a chamber. In an embodiment, the chamber presents one of a
circular cross section (e.g. a tube), a rectangular cross section
(e.g. a flat passage), and a triangular cross section. In an
embodiment, the device further comprises an opening for allowing
the biological sample to enter the chamber. This embodiment is
applicable to both fluid and solid biological samples.
[0018] In an embodiment, the device further comprises a second
opening allowing the biological sample to exit the chamber. This
embodiment is applicable to both fluid and solid biological
samples. In an embodiment, the biological sample is comprised by a
container. This embodiment is applicable to both fluid and solid
biological samples.
[0019] In an embodiment, an inner wall of the chamber is
essentially in contact with the container, when the container is
inserted into the chamber. This leads to an advantageous heating
situation since effectiveness of contact heating increases in case
there is contact between the heating means and the container
comprising the biological sample. In an embodiment, the container
is one of a needle and a tube. This embodiment is applicable to
both fluid and solid biological samples.
[0020] In an embodiment, the opening presents a seal, such as an O
ring, allowing insertion of the container comprising the biological
sample into the container. In an embodiment, the chamber comprises
a plurality of openable matching parts and a locking means for
maintaining the plurality of openable matching parts in a locked
state. By opening the matching parts and thereby allowing placing
the container, or the biological sample directly, between the
matching parts and closing, leads to an effective way of heating
the biological sample.
[0021] In an embodiment, the device further comprises a steam
generator with an inlet, wherein the chamber further comprises a
steam inlet. This allows the biological sample to be heated using
steam, which offers an effective way of heating the biological
sample. To control condensation temperature by changing pressure is
an alternative to accommodate effective heat transfer to the
biological sample.
[0022] In embodiments, the steam may be replaced by a fluid. In an
embodiment, the power transfer location is constituted by at least
one plate. In an embodiment, the heating means is one of a
radiation generator and a micro wave generator. In an embodiment,
the power transfer location presents a large power transfer
surface. In an embodiment, the power transfer location is
constituted by a warm plate receiving a biological sample or a
container comprising the biological sample. In this embodiment, the
biological sample, or the container comprising the biological
sample, is maintained in a position in which there is a contact
between the power transfer location and the biological sample, or
the container comprising the biological sample by a negative
pressure, or even vacuum, for instance acting via a hole through
the warm plate, and thus maintaining the biological sample, or the
container, in contact with the warm plate. By increasing the
pressure, the biological sample, or container, may be removed.
BRIEF DESCRIPTION OF DRAWINGS
[0023] In FIG. 1, steps of an embodiment of the method are
shown.
[0024] In FIG. 2, four schematic illustrations of preferred shapes
of biological samples are shown.
[0025] In FIG. 3, an embodiment of a device for preparing a
biological sample for biological analyses is shown.
[0026] In FIG. 4, an embodiment of a device for preparing a
biological sample for biological analyses is shown.
[0027] In FIG. 5, an embodiment of a device for preparing a
biological sample for biological analyses is shown.
[0028] In FIG. 6, an embodiment of a device for preparing a
biological sample for biological analyses is shown.
[0029] In FIG. 7, an embodiment of a device for preparing a
biological sample for biological analyses is shown.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0030] Steps of a method for preparing a biological sample for
biological analyses are shown in FIG. 1 in combination with its
application when in use. The biological sample comprises at least
one amino acid sequence from an organism. The at least one amino
acid sequence is a subject of degradation. First the biological
sample is physically prepared to enable altering a secondary
structure of the at least one amino acid sequence to a deactivated
secondary structure. In an embodiment, this step comprises shaping
the biological sample to enable altering a secondary structure of
the at least one amino acid sequence to a deactivated secondary
structure. This is followed by altering the secondary structure of
the at least one amino acid sequence to a deactivated secondary
structure by heating the biological sample.
[0031] In FIG. 2, four embodiments of shapes of biological samples
are shown. Two parallelipeds 1, 3 are shown. To the right a
cylinder shaped biological sample 5 and a biological sample 7
covered in filler 9 and provided in a paralleliped are shown. The
first three samples 1, 3, 5 are a thin and oblong in relation to
their thickness, facilitating fast heat transfer in case of contact
heat conduction by at least of one gas, condensing gas, and at
least one warm plate contacting the biological sample 1, 3, 5, or
at least contacting a container comprising the biological sample 1,
3, 5. In view of the filler 9, it presents similar electric
properties as the biological sample 7.
[0032] Embodiments of a device for preparing a biological sample
for biological analyses will now be discussed. An embodiment of a
device 11, shown in FIG. 3, comprises a chamber 13 for receiving
the biological sample 1, 3, 5, and heating means 15 for altering
the secondary structure of the at least one amino acid sequence to
a deactivated secondary structure. The embodiment also comprises an
opening 17 for allowing the biological sample 1, 3, 5 to enter the
chamber 13. The biological sample 1, 3, 5 comprised by a container
(not shown), such as a tube or a needle, is entered into the
chamber 13 via the opening 17 and an inner wall 19 of the chamber
13 is essentially in contact with the container, when the container
is inserted into the chamber 13. A thermally insulating layer 21 is
provided around the chamber 13. A heat sensor 23 and heat control
means 25 are provided for controlling the heating means 15. When
the biological sample 1, 3, 5 is in the chamber 13, the biological
sample comes in contact with the inner wall 19 of the chamber 13,
thus transferring heat to the biological sample 1, 3, 5. The heat
control means 25 may incorporate a timing function controlling the
heating means 15 so that the biological sample 1, 3, 5 is exposed
to an effective amount of heating. Also, the device 11 is powered
by a battery or means via a cord (not shown in FIG. 3). In this
embodiment, the biological sample 1, 3, 5 may be either fluid or
solid.
[0033] An embodiment of a device 31, shown in FIG. 4, comprises a
chamber 13 for receiving a fluid biological sample and heating
means 15 for altering the secondary structure of the at least one
amino acid sequence to a deactivated secondary structure. A
thermally insulating layer 21 is provided around the chamber 13. A
heat sensor 23 and heat control means 25 are provided for
controlling the heating means 15. The embodiment also comprises an
opening 17 for allowing the fluid biological sample to enter the
chamber 13. The fluid biological sample flows via the opening 17
into the chamber 13. This embodiment further comprises a second
opening 33 allowing the fluid biological sample to flow out of the
chamber 13. A heat sensor 23 and heat control means 25 are provided
for controlling the heating means 15. As the fluid biological
sample flows through the chamber 13, the fluid biological sample
comes in contact with an inner wall 19 of the chamber 13, thus
transferring heat to the fluid biological sample. The heat control
means 25 may incorporate a timing function controlling the heating
means 15. Also, the device 31 is powered by a battery or means via
a cord (not shown in FIG. 4). In this embodiment, the biological
sample is non-coagulating. The biological sample is physically
shaped during the flow in the chamber 13. The chamber 13 presents a
large heat transfer surface in relation to its volume. In this
embodiment, the cross section of the chamber 13 is designed so that
it presents a wide base and a low height, with the intention of a
fast and uniform heat transfer through the whole biological
sample.
[0034] In an embodiment of the device, shown in FIG. 5, the device
41 comprises a chamber 13 for receiving a biological sample 1, 3,
5, and heating means 25 for altering the secondary structure of the
at least one amino acid sequence to a deactivated secondary
structure. The embodiment also comprises an opening 17 for allowing
the biological sample 1, 3, 5 to enter the chamber 13. The
biological sample 1, 3, 5 comprised by a container 43, such as a
tube or a needle, is entered into the chamber 13 via the opening
17. The opening 17 presents an O ring 45, allowing sealed insertion
of the container 43 into the chamber 13. In this embodiment the
heating means 15 is constituted by a steam generator with a steam
inlet 47, wherein the chamber 13 further comprises a steam pressure
control (not shown). As the steam enters the chamber 13 the
container 43 and the biological sample 1, 3, 5 is heated. A
thermally insulating layer 21 is provided around the chamber 13. A
heat sensor 23 and heat control means 25 are provided for
controlling the heating means 15. The heat control means 25 may
incorporate a timing function controlling the heating means 15.
Also, the device 41 is powered by a battery or means via a cord
(not shown in FIG. 5). In this embodiment, the biological sample 1,
3, 5 is fluid or solid. In this embodiment there is space between
the container 43 and an inner wall of the chamber 13. This allows
fast and uniform heat transfer through the whole biological sample
1, 3, 5.
[0035] In an embodiment of the device shown in FIG. 6, the device
51 comprises a chamber 13 for receiving the biological sample 1, 3,
5, and heating means 25 for altering the secondary structure of the
at least one amino acid sequence to a deactivated secondary
structure. The chamber 13 comprises two openable matching parts 53,
55 and a locking means 57 for maintaining the two openable matching
parts 53, 55 in a locked state. The biological sample 1, 3, 5,
comprised by a container 43, such as a tube or a needle, is put
into the chamber 13 when the openable matching parts 53, 55 are in
an opened state. After closing and locking the two openable
matching parts 53, 55, the heating means 25 is activated.
Alternatively, the heating 25 is already activated. A hinge 57 is
provided along sides of the openable matching parts 53, 55. The
device 51 according to this embodiment is opened and closed as
indicated by the arrows. A thermally insulating layer 21 is
provided around the chamber 13. A heat sensor 23 and heat control
means 25 are provided for controlling the heating means 25. The
heat control means 25 may incorporate a timing function controlling
the heating means 15. Also, the device 51 is powered by a battery
or means via a cord (not shown in FIG. 6). In this embodiment, the
biological sample 1, 3, 5 is fluid or solid. In this embodiment, an
inner wall 19 of the chamber 13 is essentially in contact with the
container 43. This allows fast and uniform heat transfer through
the whole biological sample 1, 3, 5. In another embodiment, the
number of openable matching parts 53, 55 is three.
[0036] In an embodiment, the power transfer location is a single
plate heated by the heating means and the biological sample,
regardless whether placed in a container, receives heat from one
side, i.e. there is a single contact surface allowing power to
transfer into the biological sample. In an embodiment of the device
shown in FIG. 7, the device 61 comprises a chamber 13 for receiving
the biological sample 7 covered in a filler 9, and heating means 15
for altering the secondary structure of the at least one amino acid
sequence to a deactivated secondary structure. The heating means 15
is constituted by a micro wave generator.
[0037] The embodiment also comprises an opening 17, such as a door,
for allowing the biological sample 7 to enter the chamber 13. The
biological sample 7, comprised by the filler 9 presenting a similar
dielectric constant as the biological sample, is entered into the
chamber 13 via the opening 17. A heat sensor 23 and heat control
means 25 are provided for controlling the heating means 15. The
heat control means 25 may incorporate a timing function controlling
the heating means 15. Also, the device 61 is powered by means via a
cord (not shown in FIG. 7). In this embodiment, the biological
sample is fluid or solid.
[0038] While the present invention has been described in accordance
with preferred compositions and embodiments, it is to be understood
that certain substitutions and alterations may be made thereto
without departing from the spirit and scope of the following
claims.
* * * * *