CN104156739A - Electronic tag affinity ligand library and use - Google Patents
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Abstract
The invention relates to an electronic tag affinity ligand library, construction method and use. Electronic tag technology based on ligand structure information, synthesis technology of ligands on label packaging material, target biomolecule markers, and no-mark screening technology are provided, and used for quick, peculiar, high throughput screening of affinity ligands of a target biomolecule. In the electronic tag ligand library in the invention, the affinity ligands screened for the target biomolecule can be used for research and development of protein-purified affinity materials, illness target inhibitors and non-covalent binding long-acting preparation auxiliary materials of drugs.
Description
Technical field
The present invention relates to the affinity ligand of biomedicine field, be specifically related to a kind of electronic tag affinity ligand storehouse, construction method and purposes in biological medicine.
Background technology
The combination of drug molecule and biomacromolecule (as protein) is to have good biological availability, metabolic stability and hypotoxic prerequisite.Searching out to be the important starting of new drug development with the ligand molecular of bio-target molecule high affinity and selective binding (being commonly referred to affinity ligand, lead compound, lead compound), is also the basis of preparing bio-target molecule affinity purification parting material.In order to research and develop rapidly target molecule affinity ligand, many methods over nearly more than 20 years, are developed, comprise taking structure as basic MOLECULE DESIGN and with bio-target molecule screening compounds storehouse, as U.S. Patent application US20090240033A1 " Affinity Matrix Library and Its Use " has announced affinity ligand storehouse synthetic on agarose, silica gel, cellulose, glass, Toyopearl, Hydroxyethymethacrylate, Polyacrylamide, Hyper D, Stryrenedivinylbenzene or Perfluorocarbons medium.At screening affinity ligand molecule be, every kind of medium need to be filled respectively to chromatographic column, with sample one by one pillar carry out affinity chromatography operation, assessment purification effect; The sample size that screening affinity ligand needs is many, and operating process is loaded down with trivial details, the time is long, manpower and materials demand is large.Therefore, need to set up affinity ligand storehouse and screening system for bio-target molecule simple to operate, the time is short, speed is fast, high flux is high.
Summary of the invention
The present invention utilizes electronic chip as the storage of affinity ligand structures information and the electronic tag that reads, on wrapping and encapsulating material, synthesize affinity ligand storehouse, utilize target biomolecule (mark and unmarked) to sieve storehouse, select can with the affinity ligand of target molecule combination, read affinity ligand molecular structure information in chip, composition is a kind of fast, high flux affinity ligand screening system.Electronic tag is by encapsulation, as glass tube encapsulation, encapsulating material surface group is carried out to functionalization and derive, activate, directly use chemical reaction covalent coupling aglucon molecule, and/or by the molecule of the skeleton substep synthetic affinity ligand of the coupling different kinds of molecules storehouse of many avtive spots.At screening stage, utilize mark and/or the unmarked screening of natural or gene engineering expression albumen, rapid screening arrives can be in conjunction with the affinity ligand of target biomolecule.Screening process is simple to operate, quick.
Therefore, one object of the present invention is to build an electronic tag affinity ligand storehouse, and affinity ligand information is stored in the electronic tag at part place;
Another object of the present invention is to set up the method with biomolecule rapid screening electronic tag affinity ligand, hatch in target biomolecule (mark and/or unmarked) sample and affinity ligand storehouse, read target biomolecule signal on each affinity ligand label, choose the affinity ligand label that signal is the highest, by read write line direct reading ligand structure information, thereby realize affinity ligand screening quick, that high flux target biomolecule is special.
A further object of the present invention is the affinity ligand of screening native protein or gene engineering expression albumen, development affinity purification parting material and purifying process.
In the present invention; described affinity ligand storehouse be contain several functions group at the little molecule ligand compound of glass-encapsulated tube-surface chemical coupling, the ligand compound of built-in electronic tag, form multiple special space conformation, can specific binding multiple protein; comprise: taking the electronic tag glass tube with reading and writing function as matrix of packages; and functionalization is carried out in glass tube surface; and the little molecular ligand compound of coupling and the synthetic various structures of original position; form affinity ligand storehouse, can be for the protein screening of high flux, scale.
High flux, the rapid screening of the target protein affinity ligand described in the present invention, with the preparation of affinitive material, the foundation of isolation and purification method, comprise the hatching of mark and/or unmarked target protein and affinity ligand storehouse, wash, the selecting of positive label, reading of ligand structure information, and the regeneration of the wash-out of label;
The application of screening technique described in the present invention in mark or unmarked native protein or gene engineering expression protein screening, isolation and purification, to hatch with the electronic tag glass tube in a small amount of target protein sterling and affinity ligand storehouse, select adsorption target protein positive signal electron label glass tube with fluorescence signal or other non-marked signal, read the structural information of affinity ligand;
The affinity chromatography method of the target protein described in the present invention is affinity ligand information coupling and/or the synthetic corresponding affinity ligand on chromatography media obtaining according to screening, for rapid screening and the isolation and purification of albumen.
This little molecular bionics affinity ligand is more stable than bio-molecules character, and specificity and repeatability are better, and particularly an outstanding advantage is: part storehouse that can disposable synthetic a large amount of micromolecular compound, and for screening, the isolation and purification of protein.
Brief description of the drawings
Fig. 1 is the SDS-PAGE electrophoretogram of the bionical affinitive material antibody purification of embodiment tri-
Fig. 2 is the SDS-PAGE electrophoretogram of the bionical affinitive material purifying protein of embodiment tri-A
Table 1, the little molecule ligand numbering of 100 × 100 electronics (label) glass tube and attribute column (table)
The present invention is achieved by the following technical solutions, specifically comprises the steps:
Step 1: the structure in electronic tag affinity ligand storehouse: comprise the electronic tag glass tube of some and activating reagent are placed under low temperature environment and stir 1-12 hour, in course of reaction, regulate reactant liquor pH5-10 with acid or alkaline reagent, clean electronic tag glass tube 2-20 time with organic solvent or water, reject not in conjunction with activating reagent, finally electronic tag glass tube is soaked in to the aqueous solution of ethanol or the acetone of 10%-30%, low temperature is preserved stand-by.
The electronic tag of the read-write two-dimensional numerals of reaction kinetic is write and the operation of reading by the rule of experimental design, made each electronic tag glass tube there is accordingly independently numbering; According to the size of intending building part storehouse, the electronic tag glass tube of some is divided in the container of joining (body) reactant liquor of certain concentration.Be fixed in vertical rotating baking oven, adjusting temperature is 20-55 DEG C, and rotating speed is 5-100rpm, reaction 5-48 hour, and by the Ph value of acid or alkaline solution adjusting reactant liquor to pH5-9.Question response finishes rear with ethanol, acetone, and DMF (N ' dinethylformamide) or water equal solvent washing electronic tag glass tube 5-20 time, remove unconjugated free little molecular ligand.
Write, read with coupling the electronic tag glass tube of first little molecular ligand, only need to its second can reading and writing code area numeral carry out read and write operation.
Be divided in the container that includes certain density specific aglucon solution by first little molecular ligand of the coupling of some and after completing the electronic tag glass grouping of second little molecular ligand numbering, be fixed in vertical rotating baking oven, regulate oven temperature to 45-98 (85-95) DEG C, rotating speed is 10-200rpm, reaction 24-72 hour, the pH value of every 4-8 hour assaying reaction liquid in course of reaction, and be adjusted to pH5-10 with acid or alkaline solution; After finishing, question response is cooled to room temperature.With ethanol, acetone, DMF (N ' dinethylformamide) or other organic solvents or water washing electronic tag glass tube 5-20 time, remove unconjugated free little molecular ligand.Finally electronic tag glass tube is soaked in the aqueous solution of ethanol, N ' N dimethyl formamide or acetone of 5-20%, low temperature is preserved stand-by.
Step 2: in the present invention, the high flux of described protein, rapid screening, the foundation of isolation and purification method; It comprises that the scope of target protein is that mark, cold native protein or gene engineering expression albumen and coupling have different ligands electronic tag glass tube to hatch 0.1-6h at 4-70 DEG C, the unconjugated albumen of damping fluid rinsing that is 0.005-2M by the ion concentration of pH3-12, chooses 10 labels that protein signal is the strongest, reads ligand structure information.Measure on label
Embodiment
Below in conjunction with embodiment, the present invention is further elaborated, below embodiment the present invention is only described by way of example.Clearly, those of ordinary skill in the art can, in scope of the present invention and essence, carry out various accommodations and amendment to the present invention.Need to be appreciated that, this invention is intended to be encompassed in the flexible and amendment that appended claims comprises.
Embodiment mono-:
The structure in the bionical affinity ligand of high flux storehouse
On electronic label chip can reading and writing code district by the bar code writable area of 10 figure places, and 4 figure places bar code writable area composition.The bar code writable area record of first 10 figure place for this part storehouse, writes and the operation of reading with frequency read/write.Coding and information write rule: the 0th bit digital in first 10 figure place reading and writing code district represents that the structure information in structure batch the 1st little molecule ligand storehouse in part storehouse is with writing 1.To amino-compound successively according to from No. 001 initial to No. 100 numberings, while being coupled to electronic tag as first functional group, compound coding writes the first last 6-9 position, bar code writable area; In the time being coupled to electronic tag as second functional group, compound coding writes the 1-3 position of the second bar code writable area, and the 0th writes 1.
By 20000 electronic tag glass tubes (1.5mm × 7mm), the 1st writes 1, represents the 1st batch of synthetic double-basis group part.Pack in the tapered plastic bottle of 2L, add the tetraethyl orthosilicate reagent of 500ml dry toluene and 7.5mmol3-TSL 8330 and 92.5mmol, 50 DEG C of stirring reaction 24h, the cooling dereaction liquid that inclines, with countless ethanol washing 10 times, save backup with 20% ethanol.
Compound concentration is the trichloride and triazine (C of 0.5mg/ml
3n
3cl
3)-acetone soln 1000ml, be placed in-20 refrigerator precoolings after 1 hour, 10000 electronic tag glass tubes to be activated are packed in the plastic bottle containing cold acetone solution, be placed on 4 DEG C of stirrers and react after 5 minutes, by the trichloride and triazine (C of the 0.5mg/ml preparing
3n
3cl
3)-acetone soln adds in plastic bottle, makes trichloride and triazine (C
3n
3cl
3) final concentration is 0.1mg/ml, stir and measure pH after 20 minutes, and constantly use saturated Na
2cO
3solution regulates trichloride and triazine (C
3n
3cl
3)-acetone soln pH value is pH7-8, continues stirring reaction after 4.5 hours on stirrer, with distilled water washing electronic tag glass tube 10 times, obtain in 4 DEG C of refrigerators of electronic tag (glass tube) of activation, preserve stand-by.
To amino-compound successively according to from No. 001 initial to No. 100 numbering; Get 100 electronic tags, the 1001 2-4 positions that write electronic tag the first bar code writable area, then pack the high-temperature resistance plastice bottle of 100ml into, add No. 1 little molecules of ammonia based compound solution preparing, sealing was fixed in stirring reaction case, 50 DEG C of stirring reactions 24 hours, in experiment course of reaction, first about 8 hours, within every 2 hours, measure the pH value of primary first-order equation liquid, use saturated Na
2cO
3solution (reactant liquor that water is solvent) maintains pH7-10.Question response finishes to wash successively electronic tag glass tube 5-15 time with acetone, DMF (be dissolved in organic solvent containing amino little molecular ligand) or water afterwards.In washing process, every bottle of electronic tag glass tube washs separately, avoids mixing; 4 DEG C of preservations are stand-by; To all the other amino-compound numberings, write each batch of electronic tag successively, synthetic corresponding construction writes the part of electronic tag, synthesizes upper respective compound, obtains the electronic tag word bank of all compound couplings.
From the electronic tag character library of first compound of coupling, respectively get one, totally 100 electronic tags, the 001 5-7 position that writes electronic tag the first bar code writable area, then pack the high-temperature resistance plastice bottle of 100ml into, add No. 1 little molecules of ammonia based compound solution preparing, sealing is fixed in stirring reaction case, 95 DEG C of stirring reactions 24 hours, in experiment course of reaction, first about 8 hours, within every 2 hours, measure the pH value of primary first-order equation liquid, use saturated Na
2cO
3solution (reactant liquor that water is solvent) maintains pH7-10.Question response finishes to wash successively electronic tag glass tube 5-15 time with N ' dinethylformamide and water afterwards.In washing process, every bottle of electronic tag glass tube washs separately, avoids mixing; 4 DEG C of preservations are stand-by; Continue respectively to get one from the electronic tag character library of first compound of coupling, totally 100 electronic tags, write each batch of electronic tag by all the other amino-compounds numberings, synthetic respective compound, electronic tag part storehouse 1 successively.
Embodiment bis-:
The structure in the bionical affinity ligand of high flux storehouse
By 300 electronic tag glass tubes (1.5mm × 7mm), the 1st writes 2, represents the 2nd batch of synthetic single group part.Pack in the tapered plastic bottle of 500mL, add 200ml dry toluene and 7.5mmol2, the tetraethyl orthosilicate reagent of 3-epoxypropyl trimethoxy silane and 92.5mmol, 50 DEG C of stirring reaction 24h, the cooling dereaction liquid that inclines, with countless ethanol washing 10 times, saves backup with 20% ethanol.
Get 3 electronic tags at every turn, the 2-4 position of the first bar code writable area writes 001, then pack the high-temperature resistance plastice bottle of 50ml into, add No. 1 little molecules of ammonia based compound solution preparing, sealing was fixed in stirring reaction case, 60 DEG C of stirring reactions 24 hours, in experiment course of reaction, first about 8 hours, within every 2 hours, measure the pH value of primary first-order equation liquid, use saturated Na
2cO
3solution (reactant liquor that water is solvent) maintains pH7-10.Question response finishes to wash successively electronic tag glass tube 5-15 time with DMF and water afterwards.In washing process, every bottle of electronic tag glass tube washs separately, avoids mixing; 4 DEG C of preservations are stand-by; Successively all the other amino-compound numberings are write to each batch of electronic tag, synthesize the compound of corresponding construction, obtain the electronics standard configuration body storehouse 2 of all compound couplings.
Embodiment tri-:
The application of electronic tag affinity ligand storehouse in protein purification
By human serum immunoglobulin IgG 11.8mg (5ml altogether, 2.36mg/ml) with PBS (0.01M PB+0.15M NaCl, pH7.4) to be diluted to concentration be 1.2mg/ml to damping fluid, on magnetic stirring apparatus in 4 DEG C of refrigerators, stir 5～10 minutes, get FITC (fluorescein isothiocynate) 0.12mg and antibody to be marked and carry out labeled reactant: the FITC fluorescein weighing is slowly added in the antibody after dilution, in 10 minutes, all add, stir 15h, the centrifugal 10min of 12000g, remove a small amount of sediment, the antibody-solutions of mark is packed in bag filter, with 0.01M PBS (0.01M PB+0.15M NaCl, pH7.4), 4 DEG C of dialysed overnight, finally cross and filter out free fluorescein with SepHadex G-25 desalting column, collect the fluorescence antibody of mark.
By human plasma protein fraction, with 0.01M PBS (0.15M NaCl, pH7.4), to be diluted to concentration be 3mg/ml, with the albumen of the FITC mark of respective concentration according to mark IgG: human plasma=1: the potpourri of 9 ratio preparation;
From electronics standard configuration body storehouse 1, take out 200 electronic tag glass tubes, be placed in 100ml plastic bottle, add that 30ml's prepare the human plasma protein fraction containing FITC mark IgG, under 4 DEG C of conditions, 30min is hatched in slight concussion, then takes out IgG sample, add 30ml 0.01M PBS (0.01M PB+0.15M NaCl, pH7.4) level pad at room temperature slightly shakes 10min, takes out level pad, repeats this operation 10 times.Finally electronic tag glass tube is placed in luciferase target, after preservative film parcel, be placed in FLA-5100 multifunction scanner and carry out fluorescence imaging scanning (exciting light 473nm, absorb light 532nm, resolution is 100um), the fluorescent value scanning of electronic tag glass tube and data processing and analysis.
Electronic tag glass tube high fluorescent value is picked out, washing slow (rushing liquid) the concussion washing 20min that the salt ionic concentration of selecting respectively pH10 and pH3 is 1-1.5M, repeat twice of this operation, the electronic tag glass tube of collecting respectively after twice washing carries out second-order fluorescence scanning, reject the high electronic tag glass tube of fluorescent value, retain the lower electronic tag glass tube of fluorescent value, and the aglucon information of reading electronic labels glass tube, for (1000000079, 1039, ), (1000000079, 1074), (1000000027, 1049), corresponding construction is respectively undecylamine and 4-(2 aminoethyl)-benzsulfamide, Serine and 4-(2 aminoethyl)-benzsulfamide, 2, 4, the double-basis group part of 6-Triaminopyrimidine and ethylenediamine composition.In the time of follow-up synthetic checking, be labeled as respectively bionical affinity ligand 7, bionical affinity ligand 11 and bionical affinity ligand 14.
(operation is with reference to " affinity chromatography introduction " (Lip river (C.R.Lowe) work in trichloride and triazine activation for Sepharose 6B; Liu Yuxiu translates. Science Press, May nineteen eighty-three the 1st edition), measure 3 parts of each 50ml, estimate three nitrogen piperazine molal quantitys, take respectively amino-compound (R1) 4-(2 the aminoethyl)-benzsulfamide of 5 times of molar excess, 4-(2 aminoethyl)-benzsulfamide, ethylenediamine, is dissolved in the distilled water, DMF of 100ml amount, adds respectively in three nitrogen piperazine activated medias, mix at 50 DEG C of stirring reaction 24h saturated NaHCO in course of reaction
3solution maintains pH at 7-8, has reacted rear taking-up, uses successively 10 times of volume DMF (N ' N dimethyl formamide) and water washing medium.
Take respectively again amino-compound (R2) undecylamine of 5 times of molar excess, Serine, 2,4,6-Triaminopyrimidine, is dissolved in the distilled water, DMF of 100ml amount, adds respectively, coupling 4-(2 aminoethyl)-benzsulfamide, 4-(2 aminoethyl)-benzsulfamide, three nitrogen piperazine activated medias of ethylenediamine, mix, 95 DEG C of stirring reactions 24 hours, the saturated NaHCO of course of reaction
3maintain pH between 7.0-8.0, reacted rear taking-up and used successively 10 times of volume DMF (N ' N dimethyl formamide) and water washing medium, obtain bionical affinity media 7, bionical affinity media 11 and bionical affinity media 14.The ethanol that is 20% by volume fraction is preserved stand-by.
Get respectively bionical affinity media 7, bionical affinity media 11 and the each 2ml of bionical affinity media 14 load respectively in 10ml plastics chromatographic column, and at gravity column outer wall mark, with the each bionical affinity column of 10ml PB (10mM, pH7.4) damping fluid balance.By 10 times of dilution human plasma protein fraction samples of 0.01M PBS damping fluid, get 2ml and be loaded to each affinity column, with the PB solution equilibria of 20ml0.01M, finally use 50mMGly-HCl (pH3) and the 0.1N NaOH eluant solution adhesion protein of 10ml, with the detection of 12%SDS-PAG (E) gel electrophoresis and the adsorption effect of the bionical affine parting material of analysis to IgG antibody in human plasma.The purity of bionical affinitive material 7,11 and 14 specific isolation IgG purification antibody from human serum sample is respectively 95%, 91% and 86%.
Embodiment tetra-:
Get albumin A 20mg 20ml PBS (0.01M PB+0.15M NaCl, pH7.4) damping fluid and dissolve, add after FITC (fluorescein isothiocynate) 0.15mg dissolves and mix, 4 DEG C are stirred 15h; Taking-up packs in bag filter, with 0.01M PBS (0.01M PB+0.15M NaCl, pH7.4), 4 DEG C of dialysed overnight, collects fluorescence labeling albumin A.
From electronics standard configuration body storehouse 1, take out 500 electronic tag glass tubes, be placed in 200ml plastic bottle, add 50ml fluorescence labeling albumin A (0.1mg/ml), under 4 DEG C of conditions, 30min is hatched in slight concussion, then take out electronic tag glass tube 0.01M PBS (0.01M PB+0.15M NaCl, pH7.4) damping fluid washing 10 times.Finally electronic tag glass tube is placed in luciferase target, after preservative film parcel, be placed in FLA-5100 multifunction scanner and carry out fluorescence imaging scanning (exciting light 473nm, absorb light 532nm, resolution is 100um), the fluorescent value scanning of electronic tag glass tube and data processing and analysis.
Electronic tag glass tube high fluorescent value is picked out, washing 20min is swung in the washing bradyseism that the salt ionic concentration of selecting respectively pH10 and pH3 is 1-1.5M, repeat twice of this operation, the electronic tag glass tube of collecting respectively after twice collection washing carries out second-order fluorescence scanning, reject the high electronic tag glass tube of fluorescent value, retain the lower electronic tag glass tube of fluorescent value, and the aglucon information of reading electronic labels glass tube, for (1000000002, 0000), (1000000031, 0000), (1000000065, 0000), corresponding construction is respectively n-octyl amine and histamine list group part.In the time of follow-up synthetic checking, be labeled as respectively bionical affinity ligand CL3 and bionical affinity ligand CL31.
(operation is with reference to " affinity chromatography introduction " (Lip river (C.R.Lowe) work in trichloride and triazine activation for Sepharose 6B; Liu Yuxiu translates. Science Press, May nineteen eighty-three the 1st edition), measure 3 parts of each 50ml, estimate three nitrogen piperazine molal quantitys, take respectively amino-compound (R1) n-octyl amine of 5 times of molar excess, histamine and carbaniloyl hydrazine, be dissolved in 100mlDMF, add respectively in three nitrogen piperazine activated medias, mix at 50 DEG C of stirring reaction 24h saturated NaHCO in course of reaction
3solution maintains pH at 7-8, has reacted rear taking-up and has used successively 10 times of volume DMF (N ' N dimethyl formamide) and water washing medium.Obtain bionical affinity media CL3 and bionical affinity media CL31, the ethanol that is 20% by volume fraction is preserved stand-by.
Get respectively bionical affinity media CL3 and the each 2ml of bionical affinity media CL31 loads respectively in 10ml plastics chromatographic column, and at gravity column outer wall mark, with the each bionical affinity column of 10ml PB (10mM, pH7.4) damping fluid balance.Get the recombinant expressed albumin A sample of 2ml Escherichia coli fermentation (according to Chinese patent application: 201110087262.2 preparations) and be loaded to each affinity column, with the PB solution equilibria of 20ml 0.01M, finally use 50mMGly-HCl (pH3) and the 0.1N NaOH eluant solution adhesion protein of 10ml, detect and analyze the effect of bionical affine parting material purifying protein A with 12%SDS-PAG (E) gel electrophoresis.The purity of bionical affinity media CL3 and bionical affinity media CL31 purifying protein A is respectively 96%, 97%.
Table 1
| Coding 1 | Coding 2 | Chinese |
| 1000000001 | 1001 | Para aminoacetophenone |
| 1000000002 | 1002 | N-octyl amine |
| 1000000003 | 1003 | 9-aminoacridine one water hydrochloride |
| 1000000004 | 1004 | 1-amino anthraquinones |
| 1000000005 | 1005 | Two positive ethamine |
| 1000000006 | 1006 | 2-amino-Isosorbide-5-Nitrae-dicarboxylic acid (2 amino terephthalic acid (TPA)) |
| 1000000007 | 1007 | P-aminobenzoic acid |
| 1000000008 | 1008 | Hexylamine |
| 1000000009 | 1009 | N-(3-aminopropyl) imidazoles |
| 1000000010 | 1010 | The amino 1-hydrochloric acid of 1- |
| 1000000011 | 1011 | 2-aminobenzimidazole |
| 1000000012 | 1012 | Tyramine hydrochloride |
| 1000000013 | 1013 | Metaphenylene diamine hydrochloride |
| 1000000014 | 1014 | Adenine |
| 1000000015 | 1015 | Acridine yellow (nitrogen uh Huang) |
| 1000000016 | 1016 | Meta-aminophenol |
| 1000000017 | 1017 | Aminoguanidine hydrochloride glucose |
| 1000000018 | 1018 | Maloyl imines |
| 1000000019 | 1019 | CYSTINE |
| 1000000020 | 1020 | TYR |
| 1000000021 | 1021 | Trimethylamine |
| 1000000022 | 1022 | L-threonine |
| 1000000023 | 1023 | L-asparagine |
| 1000000024 | 1024 | METHIONINE |
| 1000000025 | 1025 | L-aspartic acid |
| 1000000026 | 1026 | Diaminobenzoic acid |
| 1000000027 | 1027 | 2,4,6-Triaminopyrimidine |
| 1000000028 | 1028 | 2,6-diamino-anthraquinone |
| 1000000029 | 1029 | 1,5-diamino-anthraquinone |
| 1000000030 | 1030 | The amino n-caproic acid of 6- |
| 1000000031 | 1031 | Maxamine |
| 1000000032 | 1032 | Phenyl ethylamine |
| 1000000033 | 1033 | Diamido benzo thiazole |
| 1000000034 | 1034 | 4,4 '-diaminodiphenyl ether |
| 1000000035 | 1035 | 1,2-cyclohexanediamine |
| 1000000036 | 1036 | Sulfanilic acid |
| 1000000037 | 1037 | Ortho-nitraniline |
| 1000000038 | 1038 | The amino terephthalic acid (TPA) of 2- |
| 1000000039 | 1039 | Undecylamine |
| 1000000040 | 1040 | 4-ASA sodium |
| 1000000041 | 1041 | Cyclohexylamine |
| 1000000042 | 1042 | PK-Merz |
| 1000000043 | 1043 | Allylamine |
| 1000000044 | 1044 | 1-aspartic acid |
| 1000000045 | XX045 | P-phenylenediamine (PPD) |
| 1000000046 | 1046 | O-phenylenediamine |
| 1000000047 | 1047 | Thiamine hydrochloride |
| 1000000048 | 1048 | 3-pyridyl-methanamine |
| 1000000049 | 1049 | Ethylenediamine |
| 1000000050 | 1050 | Valine |
| 1000000051 | 1051 | 1B |
| 1000000052 | 1052 | Diphenylamine |
| 1000000053 | 1053 | Pidolidone list sodium salt |
| 1000000054 | 1054 | N-phenylanthranilic acid |
| 1000000055 | 1055 | Ring imines in heptan |
| 1000000056 | 1056 | 4-aminobenzophenone |
| 1000000057 | 1057 | 5-amino-2,2,4-trimethyl-1-cyclopentyl methylamine |
| 1000000058 | 1058 | Diaminodiphenylmethane |
| 1000000059 | 1059 | Aminopyrine |
| 1000000060 | 1060 | N, N-diethyl Isosorbide-5-Nitrae phenylenediamine |
| 1000000061 | 1061 | 4-aminobenzaldehyde |
| 1000000062 | 1062 | 2-naphthylamines-1-sulfonic acid Tobias acid (tobias acid) |
| 1000000063 | 1063 | 1-amino anthraquinones |
| 1000000064 | 1064 | 4,4 diamino-diphenylamine sulfide |
| 1000000065 | 1065 | Carbaniloyl hydrazine |
| 1000000066 | 1066 | 1-amino-2-naphthol-4-sulfonic acid |
| 1000000067 | 1067 | 3-5 amido benzotrifluoride |
| 1000000068 | 1068 | DAP |
| 1000000069 | 1069 | N-butylamine |
| 1000000070 | 1070 | Triethylamine |
| 1000000071 | 1071 | Aniline |
| 1000000072 | 1072 | 3-amino, 1-propyl alcohol |
| 1000000073 | 1073 | Chaff amine |
| 1000000074 | 1074 | Serine |
| 1000000075 | 1075 | Monoethanolamine |
| 1000000076 | 1076 | 4-isopropyl aniline |
| 1000000077 | 1077 | N ' N diisopropylethylamine |
| 1000000078 | 1078 | P-phenylenediamine (PPD) |
| 1000000079 | 1079 | 4-(2 aminoethyl)-benzsulfamide |
| 1000000080 | 1080 | Benzylamine |
| 1000000081 | 1081 | Lauryl amine |
| 1000000082 | 1082 | Diethylamine hydrochloride |
| 1000000083 | 1083 | Dibenzylamine |
| 1000000084 | 1084 | Diisopropylamine |
| 1000000085 | 1085 | Di-n-butylamine |
| 1000000086 | 1086 | Di-iso-butylmanice |
| 1000000087 | 1087 | T-octanylamine |
| 1000000088 | 1088 | 3-aminoethyl pyrimidine |
| 1000000089 | 1089 | L-Histidine |
| 1000000090 | 1090 | Cyclo-dodecyl amine |
| 1000000091 | 1091 | Octadecylamine |
| 1000000092 | 1092 | L-arginine |
| 1000000093 | 1093 | The amino m-phthalic acid of 5- |
| 1000000094 | 1094 | Pidolidone sodium |
| 100000095 | 1095 | 4-amino-1,8-benzene-naphthalene diimide |
| 100000096 | 1096 | 4-morpholine propylamine |
| 1000000097 | 1097 | Amylamine, |
| 1000000098 | 1098 | 4-amino-1-naphtholate hydrochlorate |
| 1000000099 | 1099 | Paraphenetidine |
| 1000000100 | 1100 | 1-amino-5-naphthols |
Claims (10)
1. an electronic tag affinity ligand storehouse, is characterized in that the structural information of each part is stored in electronic tag, convenient can directly reading fast.
2. electronic tag affinity ligand according to claim 1 storehouse, is characterized in that electronic tag possesses information and reads, writes and erase feature, and working method is containing being undertaken by radio frequency reading and writing code device.
3. electronic tag affinity ligand according to claim 1 storehouse, is characterized in that the material package of electronic tag by organic solvent-resistant, includes but not limited to that inorganic material is if glass, macromolecular material are as polypropylene.
4. electronic tag affinity ligand according to claim 1 storehouse, directly chemistry is fixing and roll into a ball the fixing multiple compounds of skeleton substep combination by multi-active base and synthesize labyrinth part at the individualized compound of radio frequency identification tag package material surface to it is characterized in that including but not limited to organic compound cited in the present invention by described affinity ligand.
5. affinity ligand according to claim 4 storehouse synthetic method, it is characterized in that multi-active base group skeleton comprise be not limited to trichloride and triazine, Solid-phase Polypeptide is synthetic.
6. electronic tag part according to claim 1 storehouse, is characterized in that construction step includes but not limited to surface-functionalized processing, priming reaction, the reading and writing coding of electronic tag glass tube and the coupling reaction of little molecular ligand of electronic tag glass tube.
7. electronic tag affinity ligand according to claim 1 storehouse, it is characterized in that screening the affinity ligand of target molecule, when screening, target molecule can be used fluorescein, enzyme, labelled with radioisotope can be also the fusion of cold native protein or gene engineering expression.
8. electronic tag affinity ligand according to claim 1 storehouse, is characterized in that the separation and purification material of screened affinity ligand for developing goal molecule.
9. electronic tag affinity ligand according to claim 1 storehouse, is characterized in that the inhibitor of screened affinity ligand for R&D target molecule.
10. electronic tag affinity ligand according to claim 1 storehouse, is characterized in that the durative action preparation auxiliary material of screened affinity ligand for R&D target molecule.
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| CN1181720A (en) * | 1995-04-25 | 1998-05-13 | 伊萝莉公司 | Remotely programmable matrices with memories and uses thereof |
| CN101512016A (en) * | 2006-06-30 | 2009-08-19 | 埃姆比特生物科学公司 | Detectable nucleic acid tag |
| CN102719430A (en) * | 2012-06-13 | 2012-10-10 | 湖南大学 | Nucleic acid aptamer molecular beacon probe for detecting histidine-tag recombinant proteins and detection method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1181720A (en) * | 1995-04-25 | 1998-05-13 | 伊萝莉公司 | Remotely programmable matrices with memories and uses thereof |
| CN101512016A (en) * | 2006-06-30 | 2009-08-19 | 埃姆比特生物科学公司 | Detectable nucleic acid tag |
| CN102719430A (en) * | 2012-06-13 | 2012-10-10 | 湖南大学 | Nucleic acid aptamer molecular beacon probe for detecting histidine-tag recombinant proteins and detection method thereof |
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