CN104090097B - Aqueous solution of specificity binding interaction culture medium serving as binding pair - Google Patents
Aqueous solution of specificity binding interaction culture medium serving as binding pair Download PDFInfo
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- CN104090097B CN104090097B CN201410365848.4A CN201410365848A CN104090097B CN 104090097 B CN104090097 B CN 104090097B CN 201410365848 A CN201410365848 A CN 201410365848A CN 104090097 B CN104090097 B CN 104090097B
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- 239000007864 aqueous solution Substances 0.000 title claims abstract description 89
- 238000009739 binding Methods 0.000 title claims abstract description 75
- 230000027455 binding Effects 0.000 title claims abstract description 71
- 239000001963 growth medium Substances 0.000 title abstract description 21
- 230000003993 interaction Effects 0.000 title abstract description 3
- 229940126062 Compound A Drugs 0.000 claims abstract description 27
- NLDMNSXOCDLTTB-UHFFFAOYSA-N Heterophylliin A Natural products O1C2COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC2C(OC(=O)C=2C=C(O)C(O)=C(O)C=2)C(O)C1OC(=O)C1=CC(O)=C(O)C(O)=C1 NLDMNSXOCDLTTB-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000003599 detergent Substances 0.000 claims abstract description 25
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 23
- 239000001257 hydrogen Substances 0.000 claims abstract description 23
- 150000001875 compounds Chemical class 0.000 claims abstract description 20
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims abstract description 18
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 16
- 150000005846 sugar alcohols Polymers 0.000 claims abstract description 9
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims abstract description 5
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 47
- 239000000872 buffer Substances 0.000 claims description 34
- 238000006243 chemical reaction Methods 0.000 claims description 32
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 28
- 230000000694 effects Effects 0.000 claims description 25
- 239000000427 antigen Substances 0.000 claims description 20
- 102000036639 antigens Human genes 0.000 claims description 20
- 108091007433 antigens Proteins 0.000 claims description 20
- 239000011159 matrix material Substances 0.000 claims description 20
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 18
- 102000004169 proteins and genes Human genes 0.000 claims description 17
- 108090000623 proteins and genes Proteins 0.000 claims description 17
- 230000009870 specific binding Effects 0.000 claims description 17
- 235000018102 proteins Nutrition 0.000 claims description 16
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
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- 239000005977 Ethylene Substances 0.000 claims description 3
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- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 3
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 3
- 239000005018 casein Substances 0.000 claims description 3
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims description 3
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- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 3
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- 229930182478 glucoside Natural products 0.000 claims description 3
- 150000008131 glucosides Chemical class 0.000 claims description 3
- 229940092253 ovalbumin Drugs 0.000 claims description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 125000001424 substituent group Chemical group 0.000 claims description 3
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- 229940035676 analgesics Drugs 0.000 claims description 2
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- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims 1
- 125000000311 mannosyl group Chemical group C1([C@@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims 1
- IZXGZAJMDLJLMF-UHFFFAOYSA-N methylaminomethanol Chemical compound CNCO IZXGZAJMDLJLMF-UHFFFAOYSA-N 0.000 claims 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims 1
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- 239000000243 solution Substances 0.000 abstract description 17
- 230000000295 complement effect Effects 0.000 abstract description 8
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- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 7
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
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- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
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- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 2
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
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- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 102000006306 Antigen Receptors Human genes 0.000 description 1
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- 235000008434 ginseng Nutrition 0.000 description 1
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- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 1
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- 231100000989 no adverse effect Toxicity 0.000 description 1
- HEGSGKPQLMEBJL-RKQHYHRCSA-N octyl beta-D-glucopyranoside Chemical compound CCCCCCCCO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HEGSGKPQLMEBJL-RKQHYHRCSA-N 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to an aqueous solution of a specificity binding interaction culture medium serving as a binding pair. A first binding member recognizes a second complementary binding member; the solution comprises (a) a buffering solution for controlling the pH; (b) a compound A selected from the following groups: a compound defined by a general formula IR＜1＞-[[CR＜2＞R＜3＞]p-O]q-R＜4＞, polyhydric alcohols and saccharides, wherein R1 is hydrogen or a hydroxyl, R2 of each element is independent hydrogen or a hydroxyl, R3 is hydrogen, a methyl or an ethyl, R4 is hydrogen or an alkyl, p is an integer from 2 to 10, and q is an integer from 1 to 100; an additional condition is that the compound at least carries two hydroxyls; (c) a nonionic detergent.
Description
The application be the applying date on 2 26th, 2004, Application No. No.200480042127.4, entitled " one
Kind as combine to specific binding reaction culture medium aqueous solution " Chinese invention patent application divisional application.
The present invention relates to be used as with reference to specific binding reaction culture medium aqueous solution.
Background of invention
It is known to use the immunoassay that one or more antibody detects substances (analyte) in a kind of sample.
The development of immunoassay method enhances the sensitivity of the test.Although having developed in the last few years, wish to
Eliminate the impact of compound present in nonspecific association reaction, cross reaction and substrate.
Immunoassay depends on first binding members (such as a kind of antigen or part) and binding members of binding members pair
To a kind of ability that specifically binds of the second binding members (such as antibody or receptor).In order to determine the continuity of this combination, use
Detectable part labelling contains the conjugate of a side of this binding members.This binding members are to being a kind of antigen and pin
Antibody to the antigen.
Immunoassay can be carried out with emulative immunoassay format or with sandwich immunoassays form.Competing
In the immunoassay format of striving property, antigen can be fixed on solid phase material, and is attached to the detectable portion on solid phase material
The quantity divided can be detected, measure and which is associated with the quantity of antibody present in sample.The example bag of solid phase material
Include pearl, granule, microgranule etc..In sandwich immunoassays form, by the sample containing for example a kind of antibody and a hatching egg
White matter such as antigen contacts.The antigen is fixed on solid phase material.The example of solid phase material includes pearl, granule, micro-
Grain etc..The solid phase material is generally with a kind of by second antigen or antibody tormation of detectable part labelling.Then distinguish
On the solid phase material the second antigen or antibody respectively with corresponding antibodies or antigen binding, one or more washing steps with
After removing unconjugated material, a kind of indicator substances such as substance that show color is added so as to react with detectable part, produce
Detectable signal, such as color change.Then detect the color change, metering by itself and antibody levels phase present in sample
Association.In addition it is used for optimization processing microgranule, antigen, conjugate and participates in chemistry it should be noted that also needing to various diluent and buffer
Other test components of reaction.
In order to optimum is obtained in immunoassay, for (such as antibody and anti-of association reaction between binding partner
Former reaction or part and receptor generate complex) solution must provide for a kind of culture medium to optimize the energy of antibodies bind antigen
Power, or a kind of culture medium is must provide for optimize the ability of ligand binding receptor, while strong reduce even preventing non-spy
The opposite sex interacts, low affinity is combined and matrix effect, in order to avoid generation error signal.
In order to eliminate non-specific interaction and cross reaction, detergent is added after association reaction and is washed by someone
Removing non-specific binding in buffer used by step.
For immunoassay, as the analysis of the western markings, elisa (ELISA) etc., using containing benefit
Filled with bovine serum albumin and 0.01 to 0.05 (v/v)The solution of 20 phosphate buffered saline (PBS) as culture medium,
For the association reaction between binding partner (such as antibody and antigen).But it is often found that with this buffer of prior art
Cannot be avoided non-specific or low affinity combination, cross reaction and matrix effect.For example, when research one kind is related to multiple analyses
When the CRP of analyte detection is tested, ask as the cross reaction and matrix effect that bring using polynary antibody have apparently become one
Topic, which can not be solved by using conventional immunoassay buffer.
Therefore it is an object of the invention to provide a kind of solution, as the training of the specific binding reaction for binding members pair
Foster base is used, wherein strong reduce or or even prevent non-specific binding, the combination of low affinity, cross reaction and substrate effect
Should.Moreover, it is an object that a kind of method of immunity, wherein non-specific and low affinity is combined, is intersected instead
Should be lowered with matrix effect or prevent.
Summary of the invention
The purpose of the present invention is as the specific binding reaction for binding members pair by using a kind of aqueous solution
Culture medium and realize, wherein the first binding members recognize the second binding members of its complementation, the solution contains
A) control a kind of buffer of pH；
B) the compound A being selected from the group：
- by formula I R1-[[CR2R3]p-O]q-R4The compound of definition, wherein R1It is hydrogen or hydroxyl, R in each unit2It is independent
Ground is hydrogen or hydroxyl, R3It is hydrogen, methyl, ethyl, R4It is hydrogen or alkyl, p is 2 to 10 integer, and q is of 1 to 100
Integer, collateral condition are that the compound at least carries two hydroxyls；
- polyhydric alcohol；
- saccharide；
C) nonionic detergent.
In the formula I of compound A, if R4It is hydrogen, adjacent residue R2And hydrogen.If R1It is hydroxyl, adjacent is residual
Base R2It is hydrogen.In a preferred embodiment, the q in the molecular formula of compound A is an integer from 1 to 50, more
It is preferred that from 1 to 30.
Present inventors discovered unexpectedly that the present invention aqueous solution can reduce in immunoassay cross reaction, base
Mass effect, non-specific binding and low affinity are combined.Moreover, it has been found that using the thermophilic opposite sex can be prevented during aqueous solution of the present invention
The impact that brings of antibody (human anti-mouse antibody).Even if in addition, in the case where blood plasma is applied to, can be kept away with the buffer
Exempt from rheumatism (rheuma) factor, the negative effects that hemoglobin, bilirubin and triglyceride bring.
" binding members to " are including one " the first binding members " and one " the second binding members " here.Two kinds of combinations
Member can specifically bind each other.First binding members of binding members pair can be antigen or part respectively.Second is combined into
Member's (such as respectively antibody or receptor) specifically recognizes and combines the first binding members (such as respectively antigen or part).Second
Binding members are corresponding binding members, therefore also referred to as " corresponding binding members ".Technical staff can understand that term " first " is tied
Synthesis person and " second " binding members can be such as antigen and corresponding antibodies respectively, or vice versa it is as the same.
The aqueous solution of the present invention represents a kind of Generic buffer, and which is carried out in various substrate (such as blood plasma, serum etc.)
Immunoassay and association reaction in as culture medium.In the case of using multiple analyte, if for example using protein core
Piece, needs several (or multiple) analytes of synchronous incubation and several antibody, Jing that non-specific binding and cross reaction often occur.
In many cases it was observed that this association reaction for being not intended to obtain.Using standard ELISA buffer known in the art not
Can prevent this cross reaction from affecting.In addition in the prior art, compared with the method for other references, the Natural Samples for being adopted
Matrix effect can be caused, this can cause the measured value of mistake.Deposit among this term " substrate " refers to Natural Samples (such as serum)
All compounds；Term " substrate " particularly relate to organically, especially biologic artifact, such as protein.
The aqueous solution of the present invention can serve as respectively with reference to association reaction culture medium, as immunoassay and
The sample dilution buffer and antibody and the dilution buffer of antigen of association reaction.Multiple analyte immunity is can be applicable to further
Determine and protein analyses (proteomics), wherein the intersection of fluorophor traget antibody for being prevented from being not intended to obtaining is anti-
Should.Fluorophor traget antibody often combines other oroteins with non specific manner.Can be kept away using the aqueous solution of the present invention
Exempt from this impact.
In immunoassay such as ELISA and protein chip, can be shown further using the buffer of the present invention
Good effect.When with buffer of the present invention surface of the incubation with sessile antibody, the activity of the immobilized antibody has strengthened.
This causes analyte and the combination of immobilized antibody to strengthen.In a word, buffer of the present invention is except reduction non-specific signals and Fei Te
Outside the opposite sex affects, can also strengthen the specific signals for specifically binding between analyte and antibody that reacting positive effect brings.
The caused enhancing of this immobilized antibody activity provides the sensitivity of higher corresponding test.
The buffer of the present invention can be used for immunoassay ELISA, EIA, FIA, lateral streaming test (lateral-
Flow-test), protein chip, multiple analyte test, the western markings, the point marking, immunohistochemistry, receptors ligand examination
Test and immuno-PCR.
Detailed description of the invention
In a preferred embodiment of the invention, the aqueous solution is further included and immunity blocking nonspecific antibody knot
Close the protein of effective dose.This protein is preferably selected from bovine serum albumin, ovalbumin, casein, hyclone.Further
It is preferred that the protein is present in aqueous solution with the concentration of 0.1 to 2% (w/v), more preferably 0.5 to 1.5% (w/v) scope.This
Any antibody recognition that protein can not be used by immunoassay a bit.This unrecognized protein can be with immunity blocking by sample
The non-specific antibody that molecule that may be present or compound bring among product is combined.
In another specific embodiment, the aqueous solution contains selected from following salt：NaCl、KCl、NH4Cl.It is further excellent
Select the aqueous solution that there is the ionic strength of 100mM to 1.5mM, more preferably 200mM to 1M, even more preferably 200mM to 800mM,
Special more preferably 200mM to 600mM, most preferably 250mM to 500mM.Present inventors have surprisingly found that being used as association reaction
Culture medium high ionic strength of the buffer in the range of 200mM to 600mM further can reduce non-specific binding and
Cross reaction, while having no adverse effect specific binding reaction.
In specific preferred embodiment, the aqueous solution buffer selected from Tris (three (hydroxymethyl)-aminomethanes),
Pipes (piperazine -1,4- couple -2 ethane sulfonic aicd), Mes (4- morpholino ethane sulfonic acids), Hepes (4- (2- ethoxys) -1- piperazines
Piperazine-ethane sulfonic acid), phosphate buffer.
In another preferred embodiment, compound A is selected from the group：Poly alkylene glycol, polypropylene glycol, the third two
Alcohol, Polyethylene Glycol, ethylene glycol, monosaccharide, disaccharidase, trisaccharide, sucrose, mannose, trehalose, polyhydric alcohol, glycerol and its mixture.
In one preferred embodiment, the concentration of compound A is within the scope of 0.5 to 25% (v/v), preferably 2.0 to 20% (v/v),
More preferably 2.0 to 15% (v/v), and then more preferably 2.0 to 10% (v/v), even more preferably 2.0 to 7% (v/v), most preferably
About 5% (v/v).If compound A is liquid phase, the concentration is then % (v/v).If compound A is solid (such as saccharide),
The concentration should be understood % (w/v).
In another preferred embodiment, the aqueous solution contains to be washed as nonionic selected from following general formula compounds
Wash agent：
A) with substituent R1And R2(R1-Ph-R2) substituted phenyl, wherein R1It is C1-C9Alkyl, R2It is-O- [CH2-
CH2-O]a- H groups, wherein " a " is 5 to 40 integer, wherein R2Relative to R1At para-position, meta or ortho position；
b)
,
Wherein n, x, y and z are 5 to 40 integers, and R is fatty acid residue.
The further preferably non-ionic detergent is selected from：Poly- (glycol ether) m of dodecyl, wherein m be 5 to 40 it is whole
Number；1-O-n- octyl group-β-D- glycopyranoside (the pungent glucosides of n-) is (n-Octylglucoside)；Alkyl phenol it is poly- (ethylene glycol-
Ether) m, wherein m is 5 to 40 integer, preferred m=11 ()；1-O-n- dodecyl-β-D-glucopyranose
Base (1-4) α-D- glycopyranoside；Poly- (glycol ether) m of dodecyl, wherein m is 5 to 40 integer, preferred m=23 ()；Poly- (oxygen ethylene) (20)-anhydro sorbitol mono fatty acid ester, is preferably selected from poly- (oxygen ethylene) (20)-dehydration mountain
Pears alcohol monooleate (80), poly- (oxygen ethylene) (20)-Arlacel-20 (20), gather
(oxygen ethylene) (20)-Arlacel-40 (40), poly- (oxygen ethylene) (20)-anhydro sorbitol one is hard
Fat acid ester)；Octylphenol polyethylene (glycol ether) m, wherein m are 5 to 40 integers, preferred m=10 (X-100)。
In preferred embodiment, the concentration of non-ionic detergent is within the scope of 0.1 to 1.0% (v/v).It is excellent
The concentration of non-ionic detergent is selected in 0.15 to 1.0% (v/v), more preferably in the range of 0.2 to 1.0% (v/v), Jin Ergeng
It is preferred that in the range of 0.2 and 0.8% (v/v), in the range of even more preferably 0.25% to 0.6% (v/v), most preferably from about 0.25%
(v/v)。
One important feature of the present invention is the presence of compound A as claimed in claim 1 and the nonionic detergent
Agent.Relative to the concentration that solution is incubated in prior art immunoassay, described two components (wash by compound A and nonionic
Wash agent) each with higher concentration be present in the present invention aqueous solution in.Carry in another preferred embodiment
Aqueous solution is supplied, wherein the non-ionic detergent is 1 with the ratio of compound A：15 to 1：25, preferably from about 1：20.
In a particularly preferred specific embodiment, the aqueous solution contains the compound A in the range of 2 to 7% (v/v)
With the non-ionic detergent in the range of 0.2 to 0.8% (v/v).It is preferred that the aqueous solution has the ionic strength of 200mM to 1mM,
More preferably 200mM to 800mM, especially more preferably 200mM to 600mM, most preferably 250mM to 500mM.It is preferred that the aqueous solution contains
Have selected from following compounds as compound A：Poly alkylene glycol, polypropylene glycol, Propylene Glycol, Polyethylene Glycol, ethylene glycol are sweet
Oil and its mixture, are more preferably selected from following compounds：Polypropylene glycol, Propylene Glycol, Polyethylene Glycol, most preferably ethylene glycol, second
Glycol.
The aqueous solution of the present invention does not preferably contain dithiothreitol, DTT.Aqueous solution further preferably of the present invention does not contain β-sulfydryl second
Alcohol.
In another preferred embodiment, the pH of aqueous solution is adjusted in the range of 5.6 to 9.6, and preferably 6.0 arrive
In the range of 9.0, in the range of further preferred 6.5 to 8.0, in the range of most preferably 6.8 to 7.4.
The particularly preferred specific embodiment of of the aqueous solution be with reduce non-specific binding, cross reaction and
The ability that matrix interference affects.Compared with standard conditions, the aqueous solution of the present invention enables in particular to prevent KDIt is worth up to 10-7Low affinity
Property combine.The further preferably aqueous solution is prevented from KDIt is worth for 10-7The low affinity of M is combined, and compared with standard conditions, will
KDIt is worth for 10-7M to 10-8Middle affinity in the range of M is combined and reduces at least 90%.Still more preferably the aqueous solution is prevented from
KDIt is worth for 10-7The low affinity of M is combined, and compared with standard conditions, by KDIt is worth for 10-7M to 10-9Middle affinity in the range of M
With reference to reduction at least 90%.The aqueous solution that here " standard conditions " is made up of following condition is represented：50mM PBS (phosphate-buffered
Saline, pH 7.4), 150mM NaCl, 1% (w/v) BSA is (referring to table 1：List of references embodiment).With use another standard solution
The measured value of acquisition is compared, and is as a result identical, i.e., by 50mM PBS (pH7.4), 100mMNaCl, 0.05% (v/v)
The aqueous solution of Tween20 compositions.
Except reducing non-specific low affinity combination and the impact of matrix effect, the particularly preferably aqueous solution can increase
The binding activity of powerful antibody, the binding activity of preferred immobilized antibody, and the binding activity between part and receptor.Using this
The binding activity of immobilized antibody can be strengthened about 10% or more by invention solution.
The purpose of the present invention can be also solved by the concentrate of as described before aqueous solution of the present invention, preferably 2 to 10 times concentrations, more
It is preferred that 3 to 5 times of concentrations.
Further, the purpose of the present invention is solved by using aqueous solution of the present invention, as combine to combination it is anti-
The culture medium answered, wherein the first binding members specific recognition the second binding members with reference to its complementation.It is preferred that this is water-soluble
Culture medium of the liquid as antibody antigen association reaction, in a substituting specific embodiment, the aqueous solution is used as receptors ligand
The culture medium of association reaction.In other preferred embodiments, the aqueous solution is used as sample, reagent, part, receptor, anti-
The former, dilution buffer of antibody.The aqueous solution delays as washing after also preferably in immunoassay, association reaction is carried out
Rush liquid.
The present invention further provides it is a kind of combine to specific binding course of reaction in reduce non-specific binding and/
Or the method that cross reaction and/or matrix interference affect, wherein the first binding members recognize the second binding members of its complementation, should
Method includes the culture medium by the use of aqueous solution of the present invention as the specific binding reaction.
Another aspect of the present invention is that the aqueous solution of the present invention can be used as test kit component.In this term " reagent
Box " refers to various reagents and related substanceses, such as buffer, the carrier comprising immobilization binding members and required for being tested
Each reagent set.Therefore, the present invention provides a kind of detection kit by least one test analyte of immunoassay,
Wherein test analyte is the first binding members of binding members pair, and wherein first binding members specifically bind its complementation
Binding members, the test kit are included：
A) container containing aqueous solution of the present invention；
B) it is fixed with the complementary binding members thereon to capture the carrier of analyte；And
C) optionally, can the reagent of analyte that combined with complementary binding members of Immune discrimination, wherein the reagent (resists
Body) a kind of conjugated detection meanss；And
D) optionally, can react to produce the reagent of detectable response product with the detection meanss.
A kind of typical agents box for example can be used as ELISA kit for detect antibody such as in serum.In the feelings
According to the b) carrier thereon containing complementary binding members, such as virus antigen, for capturing analyte under condition.The analysis
Thing is antibody on serum.According to c), can the reagent of analyte be then a kind of anti-antibody described in Immune discrimination, which is capable of identify that institute
State captured antibody.
Realize the best mode of invention
The invention is characterised in that the aqueous solution used as the culture medium of the specific binding reaction for binding members pair,
Wherein the first binding members recognize its complementary second binding members, including：
A) control the buffer of pH；
B) selected from following compound A：By formula I R1-[[CR2R3]p-O]q-R4The compound of definition, wherein R1It is hydrogen
Or hydroxyl, R in each unit2It is independently hydrogen or hydroxyl, R3It is hydrogen, methyl, ethyl, R4It is hydrogen or alkyl, p is of 2 to 10
Integer, q are 1 to 100 integers, and collateral condition is that the compound at least carries two hydroxyls；
- polyhydric alcohol；
- saccharide；
C) nonionic detergent.
Combine to specific binding course of reaction in reduce non-specific binding and/or cross reaction and/or substrate
The inventive method of interference effect, wherein the first binding members recognize the second binding members of its complementation, i.e. for immunity survey
It is fixed, the method is characterized in that including using above-mentioned aqueous solution.
Preferably, find non-in the range of containing the compound A and 0.2 to 0.8% (v/v) in the range of 2 to 7% (v/v)
The aqueous solution of ionic detergent is useful.It is preferred that the aqueous solution has the ionic strength of 200mM to 1mM, more preferably 200mM is arrived
800mM, especially more preferably 200mM to 600mM, most preferably 250mM to 500mM.It is preferred that the aqueous solution contains selected from following
Compound is used as compound A：Poly alkylene glycol, polypropylene glycol, Propylene Glycol, Polyethylene Glycol, ethylene glycol, glycerol and its mixing
Thing, is more preferably selected from following compounds：Polypropylene glycol, Propylene Glycol, Polyethylene Glycol, most preferably ethylene glycol, ethylene glycol.
Following embodiments illustrate in greater detail the present invention.But, the embodiment is merely illustrative, and not limits this
Bright scope.
Description of the drawings
The quantity of detection antibody C6 combined with CRP in being displayed in sandwich assay by Fig. 1, under 450nm, absorbance is to CRP concentration
[ng/ml] maps.Detection antibody C6 is to enter as described in Example 1 at the standard conditions with the association reaction of protein C RP
OK, respectively using the sample buffer (referring to table 1) of the present invention.According to sample buffer I to the III of solution of the present invention well
Reduce the impact of matrix effect.Sample buffer I shows optimum sensitivity.The muting sensitivity of reference buffer is by strong
Matrix effect is caused.
Fig. 2 is displayed under the high background signal by caused by the non-specific binding of detection antibody P2 of polyclone two kinds not
With the impact of buffer, P2 non-specific binding capture antibody P3 (according to embodiment 2).Deposit without analyte in this experiment
.Compared with the high background signal with reference to embodiment buffer II, sample buffer I significantly reduces non-specific binding.
Embodiment
Embodiment 1：Reduce matrix effect
The capture antibody C2 of 100 μ l dilutions is added in each hole of microwell plate (C8 StarWell Module, NUNC) (most
1 μ g/ml of final concentration are in PBS- buffer) and covered with plate sealing gasket.Capture antibody target CRP (c- reactive proteins).Then
The plate is incubated at room temperature 5 hours.Remove sealing gasket, lavation buffer solution with 300 μ l of every hole (10mM phosphate,
7.4) 350mMNaCl, 0.05% Tween, pH wash the plate 4 times.Then (PBS- delays to add 200 μ l blocking solutions to each hole
Rush liquid, pH 7.4,1% BSA).4 DEG C of overnight incubations after being covered with plate sealing gasket.The dilution analysiss in rabbit anteserum (0-5ng/ml)
Thing CRP (c reactive proteins) is simultaneously incubated 30min at room temperature.In different sample buffers and with reference to embodiment buffer (ginseng
Detection antibody C6 (targeting CRP) of dilution biotin labeling in being shown in Table 1).Ultimate density in each prepared product is 4 μ g/ml.Contain
There is the rabbit anteserum reference material of CRP by the sample buffer containing detection antibody with 1:2 dilutions.30min is incubated at room temperature.
Remove plate sealing gasket and the plate 4 times is washed with 300 μ l lavation buffer solutions.Rap the plate so as to be dried, thoroughly remove remaining washing
Buffer.100 μ l CRP preparations are added to each hole.The plate is covered with plate sealing gasket and slight oscillatory is incubated 4h at room temperature.So
Wash the plate afterwards again.The NeutrAvidin of 100 μ l dilutions is added to each holeTM- horseradish peroxidase conjugatess (ultimate density
0.05 μ g/ml are in PBS).Slight oscillatory is incubated plate 1h at room temperature.Then the plate is washed again.By equal-volume
'sTwo kinds of solution mixing of TMB Substrat, and immediately plus 100 μ l are in each hole.It is incubated at room temperature
The plate is until there is desired color.Color turns to light blue from clarification change.In last step, 150 μ l 2M are added to each hole
H2SO4Stopped reaction, reads absorbance at 450 nm with elisa plate reader (Molecular equipment).Fig. 1 depicts difference
Impact of the buffer to matrix effect.Table 1 shows the result of the test.In table 1 non-specific binding, low affinity combine and
The reduction of matrix effect is represented with "+" in " result " hurdle.Compared with reference to embodiment buffer ("-"), the numerical statement of "+"
Show the quantity that non-specific binding, low affinity are combined and matrix effect is reduced.Fig. 1 shows detection antibody C6 combined with CRP
Quantity, at 450 nm absorbance the concentration [ng/ml] of CRP is mapped.Sample buffer I shows optimum sensitivity.Reference
The muting sensitivity of buffer is caused by strong matrix effect.
Embodiment 2：Reduce the non-specific binding of polyclone detection antibody
In ensuing experiment, in each hole of microwell plate (C8 StarWell Module, NUNC), 250 μ l are added to dilute
Capture antibody P3 (multi-clone rabbit protease inhibitor, has preparation by oneself, and ultimate density 1g/ml is in PBS- buffer) and sealed with plate
Pad is covered.The plate is incubated at room temperature 4 hours.After removing sealing gasket, lavation buffer solution with 300 μ l of every hole (10mM phosphate,
7.4) 350mM NaCl, 0.05% Tween, pH wash the plate 4 times.Then (PBS- delays to add 200ul blocking solutions to each hole
Rush liquid, pH 7.4,1% BSA).Again with 4 DEG C of overnight incubations after sealing gasket covering.Respectively with reference to embodiment buffer II or
In sample buffer I (referring to table 1), polyclone detection antibody P2 of dilution biotin labeling (have by oneself by multi-clone rabbit protease inhibitor
Preparation) (ultimate density in each preparation is 10 μ g/ml), and (250 μ l are per hole to add each hole；Five times of repetition experiment).
Plate 2h is incubated under room temperature.Remove plate sealing gasket and 4 times are washed with 300 μ l lavation buffer solutions.Rap the plate so as to be dried, it is thorough
Bottom removes remaining lavation buffer solution.The NeutrAvidin of 250 μ l dilutions is added to each holeTM- horseradish peroxidase conjugatess are (most
0.5 μ g/ml of final concentration are in PBS).Slight oscillatory is incubated plate 1h at room temperature.Then the plate is washed again.Will etc.
The Immuno of volumeTwo kinds of solution mixing of TMB Substrat, and directly plus 100 μ l are in each hole.In room temperature
Lower incubation plate is until there is desired color.Color turns to light blue from clarification change.In last step, add to each hole
150μl 2M H2SO4Stopped reaction, reads absorbance at 450 nm with elisa plate reader (Molecular equipment).Fig. 2 shows
Two kinds of different buffer under the high background signal by caused by the non-specific binding of detection antibody P2 of polyclone are shown
Affect, the P2 non-specific binding captures antibody P3.Fig. 2 shows that the sample buffer I of the present invention can reduce background signal.
Analyte is not used in this experiment.Polyclonal serum is used as antibody in addition.Polyclonal serum resists comprising many different
Body, the equal targeting target protein of these antibody.Many antibody can be combined with low or lower affinity, and some antibody then can be with
Affinity is combined, and only a kind of or minority antibody can be combined with high affinity.As shown in Fig. 2 by using the slow of the present invention
Liquid is rushed, can prevent the low affinity of corresponding antibodies from combining, affinity is combined at least reducing.Therefore, because the present invention is water-soluble
The characteristic of liquid, once a kind of analyte is added in such test, signal to noise ratio will be improved.
Table 1：The table shows the experimental result of embodiment 1：As a result (=non-specific, low affinity is combined and matrix effect
Reduction)
The present invention relates to implementation below：
Embodiment 1. it is a kind of as combine to specific binding reaction the aqueous solution that uses of culture medium, wherein
First binding members recognize the second binding members of its complementation, and the solution contains
A) control the buffer of pH；
B) the compound A being selected from the group：
- by formula I R1-[[CR2R3]p-O]q-R4The compound of definition, wherein R1It is hydrogen or hydroxyl, R in each unit2It is independent
Ground is hydrogen or hydroxyl, R3It is hydrogen, methyl, ethyl, R4It is hydrogen or alkyl, p is 2 to 10 integer, and q is 1 to 100 integer, it is subsidiary
Condition is that the compound at least carries two hydroxyls；
- polyhydric alcohol；
- saccharide；
C) nonionic detergent.
The aqueous solution of 2. embodiment 1 of embodiment, further comprising the effective of immunity blocking non-specific antibody combination
The protein of amount.
The aqueous solution of 3. embodiment 2 of embodiment, wherein the protein be selected from bovine serum albumin, ovalbumin,
Casein, hyclone.
The aqueous solution of 4. embodiment 2 or 3 of embodiment, wherein the concentration of the protein is in 0.1 to 2% (w/v) model
Within enclosing, preferably in the range of 0.5 to 1.5% (w/v).
The aqueous solution of any one of 5. embodiment 1 to 4 of embodiment, wherein the solution comprising selected from NaCl, KCl,
NH4The salt of Cl.
The aqueous solution of any one of 6. embodiment 1 to 5 of embodiment, wherein the solution has 100mM to 1.5M's
Ionic strength, preferred 200mM to 1M, more preferably 200mM to 800mM, even more preferably 200mM to 600mM, most preferably
250mM to 500mM.
The aqueous solution of any one of 7. embodiment 1 to 6 of embodiment, wherein the buffer is selected from：Tris (three (hydroxyls
Methyl)-aminomethane), Pipes (piperazine-Isosorbide-5-Nitrae-bis- -2 ethane sulfonic aicd), Mes (4- morpholino ethane sulfonic acids), Hepes (4-
(2- ethoxys) -1- piperazines-ethane sulfonic acid), phosphate buffer.
The aqueous solution of any one of 8. embodiment 1 to 7 of embodiment, wherein the compound A is selected from the group：Poly- alkylene
Base glycol, polypropylene glycol, Propylene Glycol, Polyethylene Glycol, ethylene glycol, monosaccharide, disaccharidase, trisaccharide, sucrose, mannose, trehalose are polynary
Alcohol, glycerol and its mixture.
The aqueous solution of any one of 9. embodiment 1 to 8 of embodiment, wherein the concentration of the compound A is arrived 0.5
Within the scope of 25% (v/v), preferably in the range of 2.0 to 20% (v/v), more preferably in the range of 2 to 15% (v/v), enter one
In the range of more preferably 2.0 to 10% (v/v) of step, in the range of even more preferably 2.0 to 7.0% (v/v), most preferably from about 5% (v/
v)。
The aqueous solution of any one of 10. embodiment 1 to 9 of embodiment, wherein the non-ionic detergent is formula
Compound, which is selected from
A) with substituent R1And R2(R1-Ph-R2) substituted phenyl, wherein R1It is C1-C9Alkyl, R2It is-O- [CH2-
CH2-O]a- H groups, wherein " a " is 5 to 40 integer, wherein R2Relative to R1At para-position, meta or ortho position；
b)
Wherein n, x, y and z are 5 to 40 integers, and R is fatty acid residue.
The aqueous solution of any one of 11. embodiment 1 to 9 of embodiment, the non-ionic detergent are selected from：Dodecyl
Poly- (glycol ether)m, wherein m is 5 to 40 integer；1-O-n- octyl group-β-D- glycopyranoside (the pungent glucosides of n-)；Alkyl phenol
Poly- (ethylene glycol-ether)m, wherein m is 5 to 40 integer, preferred m=11 (Nonidet)；1-O-n- dodecyl-β-D-
Glycopyranosyl (1-4) α-D- glycopyranoside；Dodecyl poly- (glycol ether)m, wherein m is 5 to 40 integer, preferred m
=23 ()；Poly- (ethylene oxide) (20)-anhydro sorbitol mono fatty acid ester, is preferably selected from poly- (ethylene oxide)
(20)-anhydro sorbitol monooleate (80), poly- (ethylene oxide) (20)-sorbitan monolaurate (20), poly- (ethylene oxide) (20)-anhydro sorbitol list Palmic acid (40), poly- (ethylene oxide)
(20)-anhydro sorbitol monostearate)；Octylphenol polyethylene (glycol ether)m, wherein m is 5 to 40 integer, preferred m=10 (X-100)。
The aqueous solution of any one of 12. embodiment 1 to 11 of embodiment, wherein the concentration of the non-ionic detergent
Within the scope of 0.1 about 1.0% (v/v), preferably in the range of 0.15 to 1.0% (v/v), more preferably in 0.2 about 1.0% (v/
V) in the range of, in the range of even more preferably 0.2 to 0.8% (v/v), even more preferably 0.25% to 0.6% (v/v) scope
It is interior, most preferably from about 0.25% (v/v).
The aqueous solution of any one of 13. embodiment 1 to 12 of embodiment, wherein the non-ionic detergent and chemical combination
The ratio of thing A is 1：15 to 1：25, preferably from about 1：20.
The aqueous solution of any one of 14. embodiment 1 to 13 of embodiment, wherein the solution does not contain two sulfur threoses
Alcohol.
The aqueous solution of any one of 15. embodiment 1 to 14 of embodiment, wherein pH are adjusted at 5.6 to 9.6 scopes
It is interior, in the range of preferably 6.0 to 9.0, in the range of more preferably 6.5 to 8.0, in the range of most preferably 6.8 to 7.4.
The aqueous solution of any one of 16. embodiment 1 to 15 of embodiment, with reducing, non-specific binding, intersection are anti-
Should and matrix interference affect ability.
The aqueous solution of any one of 17. embodiment 1 to 16 of embodiment, is prevented from KDIt is worth up to 10-7The low affinity of M
With reference to.
The aqueous solution of any one of 18. embodiment 1 to 17 of embodiment, is prevented from KDValue up to 10-7The low affinity of M
Property combine, and by KDIt is worth for 10-7M to 10-8Middle affinity in the range of M is combined and reduces at least 90%.
The aqueous solution of any one of 19. embodiment 1 to 18 of embodiment, is prevented from KDIt is worth for up to 10-7The low parent of M
Conjunction property is combined, and by KDIt is worth for 10-7M to 10-9Middle affinity in the range of M is combined and reduces at least 90%.
The aqueous solution of any one of 20. embodiment 1 to 19 of embodiment, can strengthen the binding activity (affinity of antibody
Property), the binding activity (affinity) of preferred immobilized antibody.
The concentrate of the aqueous solution of any one of 21. embodiment 1 to 20 of embodiment, preferably 2 to 10 times concentrate, more
It is preferred that 3 to 5 times of concentrate.
The aqueous solution of any one of 22. embodiment 1 to 20 of embodiment as combine to association reaction training
The purposes of foster base, wherein the first binding members specific recognition the second binding members with reference to its complementation.
The aqueous solution of any one of 23. embodiment 1 to 20 of embodiment is used as the culture medium of antibody antigen association reaction
Purposes.
The aqueous solution of any one of 24. embodiment 1 to 20 of embodiment is used as the culture medium of analgesics screening platform reaction
Purposes.
The aqueous solution of any one of 25. embodiment 1 to 20 of embodiment be used as sample and the preferred part of reagent, receptor,
Antigen, the dilution buffer of antibody or the purposes as lavation buffer solution.
Embodiment 26. it is a kind of combine to specific binding course of reaction in reduce non-specific binding and/or friendship
The method that fork reaction and/or matrix interference affect, wherein the first binding members recognize the second binding members of its complementation, the side
Method includes the culture medium that the aqueous solution by the use of embodiment 1 to 20 is reacted as the specific binding.
A kind of detection kit by least one test analyte of immunoassay of embodiment 27, wherein analysis to be measured
Thing is the first binding members of binding members pair, and wherein first binding members specifically bind its complementary binding members, should
Test kit is included：
A) container of the buffer of any one containing embodiment 1 to 20；
B) it is fixed with the complementary binding members thereon to capture the carrier of analyte；And
C) optionally, can the reagent of analyte that combined with complementary binding members of Immune discrimination, wherein the reagent yoke
Unification kind detection meanss；
And d) optionally, can react to produce the reagent of detectable response product with the detection meanss.
Claims (37)
1. it is a kind of combine to specific binding course of reaction in reduce matrix effect or reduce or prevent because thermophilic different
The in vitro method of the effect that property antibody causes, wherein the first binding members recognize the second binding members of its complementation, wherein described
Specific binding reaction be to carry out in the substrate of blood plasma or serum, methods described includes using aqueous solution as the spy
The medium of anisogamy reaction, the aqueous solution contain
A) control the buffer agent of pH；
B) the compound A being selected from the group：
- polyhydric alcohol；
- saccharide；
C) non-ionic detergent,
Wherein described aqueous solution has the ionic strength of 200mM-1M, and the aqueous solution does not contain dithiothreitol, DTT.
2. it is a kind of combine to specific binding course of reaction in reduce matrix effect or reduce or prevent because thermophilic different
The in vitro method of the effect that property antibody causes, wherein the first binding members recognize the second binding members of its complementation, wherein described
Specific binding reaction be to carry out in the substrate of blood plasma or serum, methods described includes using aqueous solution as the spy
The medium of anisogamy reaction, the aqueous solution contain
A) control the buffer agent of pH；
B) following compound A：
- by formula I R1-[[CR2R3]p-O]q-R4The compound of definition, wherein R1It is hydrogen or hydroxyl, R in each unit2It is independently
Hydrogen or hydroxyl, R3It is hydrogen, methyl, ethyl, R4It is hydrogen, p is 2 to 10 integer, and q is 1 to 50 integer, and collateral condition is the change
Compound at least carries two hydroxyls；
C) non-ionic detergent,
Wherein described aqueous solution has the ionic strength of 200mM-1M, and the aqueous solution does not contain dithiothreitol, DTT.
3. the method for claim 1 or 2, wherein the effective dose that described aqueous solution is combined comprising immunity blocking non-specific antibody
Protein.
4. the method for claim 3, wherein the protein is selected from bovine serum albumin, ovalbumin and casein.
5. the method for claim 3, wherein the concentration of the protein is within the scope of 0.1 to 2w/v%.
6. the method for claim 1 or 2, wherein the effective dose that described aqueous solution is combined comprising immunity blocking non-specific antibody
Hyclone.
7. the method for claim 6, wherein the concentration of hyclone is within the scope of 0.1 to 2w/v%.
8. the method for claim 1 or 2, wherein described aqueous solution is comprising selected from NaCl, KCl, NH4The salt of Cl.
9. the method for claim 1 or 2, wherein the aqueous solution has the ionic strength of 200-800mM.
10. the method for claim 1 or 2, wherein the aqueous solution has the ionic strength of 250-800mM.
The method of 11. claim 1 or 2, wherein the buffer agent is selected from：Three (hydroxymethyl)-aminomethane, piperazine-Isosorbide-5-Nitrae-bis--
2 ethane sulfonic aicd, 4- morpholino ethane sulfonic acids, 4- (2- ethoxys) -1- piperazines-ethane sulfonic acid, phosphate buffer.
12. the method for claim 1 wherein that the compound A is selected from the group：Monosaccharide, disaccharidase, trisaccharide, polyhydric alcohol, and its mixing
Thing.
The method of 13. claim 12, wherein the polyhydric alcohol is poly alkylene glycol.
The method of 14. claim 13, wherein the poly alkylene glycol is selected from polypropylene glycol and Polyethylene Glycol.
The method of 15. claim 12, wherein the monosaccharide is mannose.
The method of 16. claim 12, wherein the disaccharidase is selected from sucrose and trehalose.
The method of 17. claim 12, wherein the polyhydric alcohol is selected from Propylene Glycol, ethylene glycol and glycerol.
The method of 18. claim 1 or 2, wherein the concentration of the compound A is within the scope of 2.0 to 20w/v or v/v%.
The method of 19. claim 1 or 2, wherein the concentration of the compound A is within the scope of 2.0 to 15w/v or v/v%.
The method of 20. claim 1 or 2, wherein the concentration of the compound A is within the scope of 2.0 to 10w/v or v/v%.
The method of 21. claim 1 or 2, wherein the concentration of the compound A is within the scope of 2.0 to 7w/v or v/v%.
The method of 22. claim 1 or 2, wherein the non-ionic detergent is general formula compound, which is selected from
A) with substituent R1And R2Substituted phenyl (R1-Ph-R2), wherein R1It is C1-C9Alkyl, R2It is-O- [CH2-CH2-
O]a- H groups, wherein " a " is 5 to 40 integer, wherein R2Relative to R1At para-position, meta or ortho position；
b)
Wherein n, x, y and z are 5 to 40 integers, and R is fatty acid residue.
The method of 23. claim 1 or 2, wherein the non-ionic detergent is selected from：Dodecyl poly- (glycol ether)m, its
Middle m is 5 to 40 integer；1-O- n-octyl-β-D- glycopyranoside (just pungent glucoside)；Alkyl phenol poly- (ethylene glycol-ether)m,
Wherein m is 5 to 40 integer；1-O-dodecyl-β-D-glycopyranosyl (1-4) α-D- glycopyranoside；Dodecyl
Poly- (glycol ether)m, wherein m is 5 to 40 integer；Poly- (ethylene oxide) (20)-anhydro sorbitol mono fatty acid ester；Octyl group
Phenol poly- (glycol ether)m, wherein m is 5 to 40 integer.
The method of 24. claim 1 or 2, wherein the concentration of the non-ionic detergent is within the scope of 0.1-1.0v/v%.
The method of 25. claim 1 or 2, wherein the concentration of the non-ionic detergent 0.15-1.0v/v% scopes it
It is interior.
The method of 26. claim 1 or 2, wherein the concentration of the non-ionic detergent is within the scope of 0.2-1.0v/v%.
The method of 27. claim 1 or 2, wherein the concentration of the non-ionic detergent is within the scope of 0.2-0.8v/v%.
The method of 28. claim 1 or 2, wherein the non-ionic detergent is 1 with the ratio of compound A：15 to 1：25.
The method of 29. claim 1 or 2, wherein pH are adjusted in the range of 5.6 to 9.6.
The method of 30. claim 1 or 2, wherein the aqueous solution is prevented from KDValue up to 10-7The low affinity of M is combined.
The method of 31. claim 1 or 2, wherein the aqueous solution is prevented from KDValue up to 10-7The low affinity of M is combined, and
By KDIt is worth for 10-7M to 10-8Middle affinity in the range of M is combined and reduces at least 90%.
The method of 32. claim 1 or 2, wherein the aqueous solution is prevented from KDValue up to 10-7The low affinity of M is combined, and
By KDIt is worth for 10-7M to 10-9Middle affinity in the range of M is combined and reduces at least 90%.
The method of 33. claim 1 or 2, wherein the aqueous solution can strengthen the binding activity of antibody.
The method of 34. claim 1 or 2, wherein the association reaction is antibody antigen association reaction.
The method of 35. claim 1 or 2, wherein the association reaction is analgesics screening platform reaction.
The method of 36. claim 1 or 2, wherein the aqueous solution is used as the dilution buffer of sample and reagent or is used as washing
Buffer.
The method of 37. claim 1 or 2, wherein the heterophil antibody is human anti-mouse antibody.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410365848.4A CN104090097B (en) | 2004-02-26 | 2004-02-26 | Aqueous solution of specificity binding interaction culture medium serving as binding pair |
| HK14112246.1A HK1198778A1 (en) | 2004-02-26 | 2014-12-04 | An aqueous solution for use as medium for the specific binding reaction of a binding pair |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200480042127.4A CN1922487A (en) | 2004-02-26 | 2004-02-26 | Water solution of specificity binding reaction culture medium used as binding pair |
| CN201410365848.4A CN104090097B (en) | 2004-02-26 | 2004-02-26 | Aqueous solution of specificity binding interaction culture medium serving as binding pair |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN200480042127.4A Division CN1922487A (en) | 2004-02-26 | 2004-02-26 | Water solution of specificity binding reaction culture medium used as binding pair |
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| CN104090097B true CN104090097B (en) | 2017-04-12 |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5616460A (en) * | 1995-06-07 | 1997-04-01 | Abbott Laboratories | Buffer composition for reagents for immunoassay |
| US6171801B1 (en) * | 1996-07-18 | 2001-01-09 | Dade Behring Marburg Gmbh | Methods for releasing a ligand from a complex |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6231982B1 (en) * | 1997-12-10 | 2001-05-15 | Dade Behring Inc. | Particle reagents having reduced matrix effects and containing an aldehyde-reactive functional group |
| DE60035127T2 (en) * | 2000-03-06 | 2008-02-14 | Dade Behring Marburg Gmbh | POLYSACCHARIDE-COATED CARRIER, THEIR MANUFACTURE AND USE |
-
2004
- 2004-02-26 CN CN201410365848.4A patent/CN104090097B/en not_active Expired - Lifetime
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2014
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5616460A (en) * | 1995-06-07 | 1997-04-01 | Abbott Laboratories | Buffer composition for reagents for immunoassay |
| US6171801B1 (en) * | 1996-07-18 | 2001-01-09 | Dade Behring Marburg Gmbh | Methods for releasing a ligand from a complex |
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|---|---|
| HK1198778A1 (en) | 2015-06-05 |
| CN104090097A (en) | 2014-10-08 |
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