CN105669861A - Method for preparing goat hair detection antibody by feature diagnosis sequence - Google Patents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
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- 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
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- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
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- G01N2333/4742—Keratin; Cytokeratin
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Abstract
The invention relates to the technical field of cultural relics detection, and discloses a method for preparing a goat hair detection antibody by a feature diagnosis sequence. The method comprises the following steps of a, synthesizing a feature diagnosis sequence; b, coupling the feature diagnosis sequence and a vector; c, performing primary immunization; d, preparing antiserums; e, purifying the goat hair detection antibody. The goat hair detection antibody prepared by the method has the advantages of high sensitivity and high specificity.
Description
Technical field
The present invention relates to historical relic detection technique field, particularly relate to a kind of method utilizing feature diagnosing sequence to prepare goathair detection antibody.
Background technology
Chinese history is long, and being unearthed from the grave that China is early stage has a large amount of goathair goods, and these, goathair goods contained a large amount of archaeological informations in ancient times.
Goathair is a kind of natural animal hair fibre. Goathair is made up of epidermis scales layer and lamina corticalis two portions, being a kind of thiozell, wherein Keratin sulfate is formed primarily of 18 kinds of a-amino acids, and is coupled to helical long chain molecule, containing carboxyl, hydroxyl and amido etc. on it, at intermolecular formation hydrogen bond and salt bridged bond etc.
Usually to the qualification of goathair goods mainly to the qualification of goathair Keratin sulfate. Goathair Keratin sulfate is a kind of hardening, and not soluble protein, the discriminating of goathair goods is caused certain difficulty by this. At present, protein detection technology is such as the modern technique that the enzyme linked immunological technology combined based on Ag-Ab is a kind of widespread use, but the antibody preparation possessing high sensitivity and specificity is a big difficult problem.
Summary of the invention
In order to solve the problems of the technologies described above, the present invention provides a kind of method utilizing feature diagnosing sequence to prepare goathair detection antibody. Goathair detection antibody prepared by the inventive method has high sensitivity and high specific.
The concrete technical scheme of the present invention is: a kind of method utilizing feature diagnosing sequence to prepare goathair detection antibody, adopts following steps:
A. the synthesis of feature diagnosing sequence: adopt Fmoc polypeptide solid-state reaction method synthesis feature diagnosing sequence, and increase cysteine residues for linked reaction at the C end of described feature diagnosing sequence.
B. the coupling of feature diagnosing sequence and carrier: select hemocyanin as carrier proteins, connects method with SPDP and described feature diagnosing sequence and described hemocyanin carry out coupling obtains complete antigen.
C. initial immunity: the complete antigen that step b is obtained is diluted with physiological saline, by the complete antigen after dilution and complete Freund's adjuvant 1:(0.8-1.1 by volume) mix, then add penicillin and Streptomycin sulphate carries out emulsification, obtain initial immunity antigen emulsion; Use initial immunity antigen emulsion that rabbit is carried out subcutaneous multi-point injection, carry out initial immunity.
D. antiserum prepare: the complete antigen obtained by step b with physiological saline dilutes, by the complete antigen after dilution and incomplete Freund's adjuvant 1:(0.8-1.1 by volume) mix, then add penicillin and Streptomycin sulphate emulsification, obtain strong immunizing antigen emulsion; After initial immunity the 3rd week and when the 5th week, use strong immunizing antigen emulsion that rabbit is carried out subcutaneous multi-point injection respectively, carry out booster immunization;After carrying out initial immunity the 7th week and when the 9th week, repeat booster immunization, when the antibody titer in rabbit blood sample reaches 1:20000, collect rabbit blood, and be separated obtained antiserum(antisera).
E. the purifying of goathair detection antibody: albumin A filler is mixed with phosphate buffer soln, vacuumizes 20-25min after stirring, obtain the albumin A filler liquid of bubble-free; Albumin A filler liquid is joined in glass column under air bubble free conditions, obtains albumen post; Albumen post is cleaned; Above-mentioned obtained antiserum(antisera) is diluted, joins after dilution and albumen post carries out loading, repeat loading once; Albumen post is cleaned again, carries out wash-out with glycine solution antagonist, after wash-out, clean albumen post, obtained antibody.
Antibody prepared by the present invention has stronger specificity, and this antibody can have obvious immune response with goathair Keratin sulfate, highly sensitive, it is possible to the goathair keratin molecule ruptured in goathair goods ancient times makes detection.
As preferably, the feature diagnosing sequence described in step a is the aminoacid sequence PVSCEATICE as shown in sequence table SEQ IDNo.1.
As preferably, the concentration of the complete antigen after diluting in step c is 0.8-1.2mg/mL.
As preferably, the concentration of the complete antigen after steps d dilution is 0.25-0.75mg/mL.
As preferably, when carrying out initial immunity, booster immunization, the injection volume of each injection point is 100 μ L.
As preferably, in steps d, being separated the method obtaining antiserum(antisera) from blood is: the blood collected is placed in set at room temperature, then solidificating blood is placed in the warm case of 36-38 DEG C and places 25-35min, the refrigerator 11-13h being placed in 4 DEG C again, makes clot fully shrink and obtain the thick antiserum(antisera) that precipitates out completely; Collecting thick antiserum(antisera), in 4 DEG C of environment, when 3000g, centrifugal 14-16min, gets supernatant liquor, is positioned over-80 DEG C of storages for subsequent use.
As preferably, the detailed process of step e is: mixed by the proportioning of 1.5g/ (6-7) mL with phosphate buffer soln by albumin A filler, vacuumize 20-25min after stirring, obtain the albumin A filler liquid of bubble-free; Then the albumin A filler liquid of 6-7 parts by volume is joined in glass column under air bubble free conditions, obtain albumen post; Being cleaned by albumen post with the phosphate buffer soln of 20 parts by volume, cleaning flow velocity is 60mL/h; With phosphate buffer soln, the antiserum(antisera) of 15 parts by volume is diluted to 20 parts by volume, joins after dilution and albumen post carries out loading, repeat loading once; Again being cleaned by albumen post with the phosphate buffer soln of 30 parts by volume, cleaning flow velocity is 60mL/h; With concentration to be 0.2mol/L, pH be 3.0 glycine solution antagonist carry out wash-out; Wash albumen post with phosphate buffer soln after wash-out, obtained antibody, antibody is loaded in centrifuge tube and stand-by in 4 DEG C of preservations.
As preferably, the concentration of phosphate buffer soln used in step e is 0.05mol/L, pH is 7.4.
It is compared with the prior art, the invention has the beneficial effects as follows:
(1) utilize feature diagnosing sequence to prepare goathair detection antibody and there is the features such as highly sensitive, high specificity, it is possible to targets identification Goral Horn albumen, it is possible to be applied to MBP enzyme linked immuno-adsorbent assay, immunofluorescence technique etc.
(2) adopt feature diagnosing sequence to prepare goathair detection antibody and the detection of goathair fabric in early stage grave can be provided a kind of responsive, special, discrimination method efficiently, for goathair archaeology and a source problem provide new scientific basis.
Embodiment
Below in conjunction with embodiment, the invention will be further described.In the following example, the coupling of feature diagnosing sequence and synthesis and feature diagnosing sequence and carrier all entrusts Huaan, Hangzhou Bioisystech Co., Ltd to complete.
Embodiment 1
Utilize feature diagnosing sequence to prepare a method for goathair detection antibody, adopt following steps:
A. the synthesis of feature diagnosing sequence: adopt Fmoc polypeptide solid-state reaction method synthesis feature diagnosing sequence, and increase cysteine residues for linked reaction at the C end of described feature diagnosing sequence. Aminoacid sequence such as the sequence table SEQ IDNo.1 of described feature diagnosing sequence show PVSCEATICE.
B. the coupling of feature diagnosing sequence and carrier: select hemocyanin as carrier proteins, connects method with SPDP and described feature diagnosing sequence and described hemocyanin carry out coupling obtains complete antigen.
C. initial immunity: with physiological saline by complete antigen weaker concn obtained for step b to 1mg/mL, by complete antigen and the complete Freund's adjuvant after dilution by volume 1:0.95 mix, then add penicillin and Streptomycin sulphate carries out emulsification, obtain initial immunity antigen emulsion; Use initial immunity antigen emulsion that rabbit is carried out subcutaneous multi-point injection, carry out initial immunity. When carrying out initial immunity, the injection volume of each injection point is 100 μ L.
D. antiserum prepare: the complete antigen weaker concn obtained by step b with physiological saline is to 0.5mg/mL, by complete antigen and the incomplete Freund's adjuvant after dilution by volume 1:0.95 mix, then add penicillin and Streptomycin sulphate emulsification, obtain strong immunizing antigen emulsion; After initial immunity the 3rd week and when the 5th week, use strong immunizing antigen emulsion that rabbit is carried out subcutaneous multi-point injection respectively, carry out booster immunization; After carrying out initial immunity the 7th week and when the 9th week, repeating booster immunization, when carrying out booster immunization, the injection volume of each injection point is 100 μ L.
When the antibody titer in rabbit blood sample reaches 1:20000, collect rabbit blood, the blood collected is placed in set at room temperature, then solidificating blood is placed in the warm case of 37 DEG C and places 30min, the refrigerator 12h being placed in 4 DEG C again, makes clot fully shrink and obtain the thick antiserum(antisera) that precipitates out completely; Collecting thick antiserum(antisera), in 4 DEG C of environment, when 3000g, centrifugal 15min, gets supernatant liquor, and obtained antiserum(antisera), is positioned over-80 DEG C of storages for subsequent use.
E. the purifying of goathair detection antibody: albumin A filler is mixed by the proportioning of 1.5g/6.5mL with phosphate buffer soln, vacuumizes 22.5min after stirring, obtain the albumin A filler liquid of bubble-free; Then the albumin A filler liquid of 6.5mL is joined in glass column under air bubble free conditions, obtain albumen post; Being cleaned by albumen post with the phosphate buffer soln of 20mL, cleaning flow velocity is 60mL/h; With phosphate buffer soln, the antiserum(antisera) of 15mL is diluted to 20mL, joins after dilution and albumen post carries out loading, repeat loading once; Again being cleaned by albumen post with the phosphate buffer soln of 30mL, cleaning flow velocity is 60mL/h; With concentration to be 0.2mol/L, pH be 3.0 glycine solution antagonist carry out wash-out; Wash albumen post with phosphate buffer soln after wash-out, obtained antibody, antibody is loaded in centrifuge tube and stand-by in 4 DEG C of preservations. The concentration of above-mentioned phosphate buffer soln is 0.05mol/L, pH is 7.4.
F. specific detection: get goathair Keratin sulfate mass concentration be 1% the bovine serum albumin solution concentration that is mixed with 1 μ g/mL be coated in enzyme plate 12h, wash three times with the phosphate buffer soln of pH7.4;Every hole adds 200 μ L, and mass concentration is bovine serum albumin solution closed 2h at 37 DEG C of 1%; 3 times are washed with the phosphate buffer soln of pH7.4, then goathair is detected the bovine serum albumin solution dilution 1:240000 that antibody by mass concentration is 1% respectively, 1:60000,1:20000,1:10000,1:5000, doubly, every hole adds 100 μ L goathair detection antibody to 1:1000,1:200, hatch 1h at 37 DEG C, wash 5 times with the phosphate buffer soln of pH7.4; The horseradish peroxidase mark two that every hole adds 100 μ L resists, and hatches 1h at 37 DEG C, described two anti-employing mass concentrations be 1% bovine serum albumin dilute 5000 times; Last every hole add the mass concentration of 100 μ L be 1% tetramethyl biphenyl amine aqueous solution in the colour developing of dark place, after 10min, add the H of 100 μ L2mol/L2SO4Termination reaction; OD is measured by microplate reader450nm, according to OD450nmValue, judges situations such as the associativities of antigen-antibody; Just can determine when OD value reaches positive criteria in sample containing Goral Horn albumen; If containing Goral Horn albumen in sample, then OD value will lower than positive criteria, according to this standard, it can be determined that the existence of goathair.
Embodiment 2
Utilize feature diagnosing sequence to prepare a method for goathair detection antibody, adopt following steps:
A. the synthesis of feature diagnosing sequence: adopt Fmoc polypeptide solid-state reaction method synthesis feature diagnosing sequence, and increase cysteine residues for linked reaction at the C end of described feature diagnosing sequence. Aminoacid sequence such as the sequence table SEQ IDNo.1 of described feature diagnosing sequence show PVSCEATICE.
B. the coupling of feature diagnosing sequence and carrier: select hemocyanin as carrier proteins, connects method with SPDP and described feature diagnosing sequence and described hemocyanin carry out coupling obtains complete antigen.
C. initial immunity: with physiological saline by complete antigen weaker concn obtained for step b to 0.8mg/mL, by complete antigen and the complete Freund's adjuvant after dilution by volume 1:0.8 mix, then add penicillin and Streptomycin sulphate carries out emulsification, obtain initial immunity antigen emulsion; Use initial immunity antigen emulsion that rabbit is carried out subcutaneous multi-point injection, carry out initial immunity. When carrying out initial immunity, the injection volume of each injection point is 100 μ L.
D. antiserum(antisera) and preparation: the complete antigen weaker concn obtained by step b with physiological saline is to 0.25mg/mL, by complete antigen and the incomplete Freund's adjuvant after dilution by volume 1:0.8 mix, then add penicillin and Streptomycin sulphate emulsification, obtain strong immunizing antigen emulsion; After initial immunity the 3rd week and when the 5th week, use strong immunizing antigen emulsion that rabbit is carried out subcutaneous multi-point injection respectively, carry out booster immunization; After carrying out initial immunity the 7th week and when the 9th week, repeating booster immunization, when carrying out booster immunization, the injection volume of each injection point is 100 μ L.
When the antibody titer in rabbit blood sample reaches 1:20000, collect rabbit blood, the blood collected is placed in set at room temperature, then solidificating blood is placed in the warm case of 36 DEG C and places 25min, the refrigerator 11h being placed in 4 DEG C again, makes clot fully shrink and obtain the thick antiserum(antisera) that precipitates out completely; Collecting thick antiserum(antisera), in 4 DEG C of environment, when 3000g, centrifugal 14min, gets supernatant liquor, and obtained antiserum(antisera), is positioned over-80 DEG C of storages for subsequent use.
E. the purifying of goathair detection antibody: albumin A filler is mixed by the proportioning of 1.5g/6mL with phosphate buffer soln, vacuumizes 20min after stirring, obtain the albumin A filler liquid of bubble-free;Then the albumin A filler liquid of 6mL is joined in glass column under air bubble free conditions, obtain albumen post; Being cleaned by albumen post with the phosphate buffer soln of 20mL, cleaning flow velocity is 60mL/h; With phosphate buffer soln, the antiserum(antisera) of 15mL is diluted to 20mL, joins after dilution and albumen post carries out loading, repeat loading once; Again being cleaned by albumen post with the phosphate buffer soln of 30mL, cleaning flow velocity is 60mL/h; With concentration to be 0.2mol/L, pH be 3.0 glycine solution antagonist carry out wash-out; Wash albumen post with phosphate buffer soln after wash-out, obtained antibody, antibody is loaded in centrifuge tube and stand-by in 4 DEG C of preservations. The concentration of above-mentioned phosphate buffer soln is 0.05mol/L, pH is 7.4.
Embodiment 3
Utilize feature diagnosing sequence to prepare a method for goathair detection antibody, adopt following steps:
A. the synthesis of feature diagnosing sequence: adopt Fmoc polypeptide solid-state reaction method synthesis feature diagnosing sequence, and increase cysteine residues for linked reaction at the C end of described feature diagnosing sequence. Aminoacid sequence such as the sequence table SEQ IDNo.1 of described feature diagnosing sequence show PVSCEATICE.
B. the coupling of feature diagnosing sequence and carrier: select hemocyanin as carrier proteins, connects method with SPDP and described feature diagnosing sequence and described hemocyanin carry out coupling obtains complete antigen.
C. initial immunity: with physiological saline by complete antigen weaker concn obtained for step b to 1.2mg/mL, by complete antigen and the complete Freund's adjuvant after dilution by volume 1:1.1 mix, then add penicillin and Streptomycin sulphate carries out emulsification, obtain initial immunity antigen emulsion; Use initial immunity antigen emulsion that rabbit is carried out subcutaneous multi-point injection, carry out initial immunity. When carrying out initial immunity, the injection volume of each injection point is 100 μ L.
D. antiserum prepare: the complete antigen weaker concn obtained by step b with physiological saline is to 0.75mg/mL, by complete antigen and the incomplete Freund's adjuvant after dilution by volume 1:1.1 mix, then add penicillin and Streptomycin sulphate emulsification, obtain strong immunizing antigen emulsion; After initial immunity the 3rd week and when the 5th week, use strong immunizing antigen emulsion that rabbit is carried out subcutaneous multi-point injection respectively, carry out booster immunization; After carrying out initial immunity the 7th week and when the 9th week, repeating booster immunization, when carrying out booster immunization, the injection volume of each injection point is 100 μ L.
When the antibody titer in rabbit blood sample reaches 1:20000, collect rabbit blood, the blood collected is placed in set at room temperature, then solidificating blood is placed in the warm case of 38 DEG C and places 35min, the refrigerator 13h being placed in 4 DEG C again, makes clot fully shrink and obtain the thick antiserum(antisera) that precipitates out completely; Collecting thick antiserum(antisera), in 4 DEG C of environment, when 3000g, centrifugal 16min, gets supernatant liquor, and obtained antiserum(antisera), is positioned over-80 DEG C of storages for subsequent use.
E. the purifying of goathair detection antibody: albumin A filler is mixed by the proportioning of 1.5g/7mL with phosphate buffer soln, vacuumizes 25min after stirring, obtain the albumin A filler liquid of bubble-free; Then the albumin A filler liquid of 7mL is joined in glass column under air bubble free conditions, obtain albumen post; Being cleaned by albumen post with the phosphate buffer soln of 20mL, cleaning flow velocity is 60mL/h; With phosphate buffer soln, the antiserum(antisera) of 15mL is diluted to 20mL, joins after dilution and albumen post carries out loading, repeat loading once;Again being cleaned by albumen post with the phosphate buffer soln of 30mL, cleaning flow velocity is 60mL/h; With concentration to be 0.2mol/L, pH be 3.0 glycine solution antagonist carry out wash-out; Wash albumen post with phosphate buffer soln after wash-out, obtained antibody, antibody is loaded in centrifuge tube and stand-by in 4 DEG C of preservations. The concentration of above-mentioned phosphate buffer soln is 0.05mol/L, pH is 7.4.
Raw materials used, equipment in the present invention, unless otherwise noted, is conventional raw material, the equipment of this area; Method therefor in the present invention, unless otherwise noted, is the ordinary method of this area.
The above; it it is only the better embodiment of the present invention; not the present invention being imposed any restrictions, every any simple modification, change and equivalent transformation above embodiment done according to the technology of the present invention essence, all still belongs to the protection domain of technical solution of the present invention.
Claims (8)
1. one kind utilize feature diagnosing sequence prepare goathair detection antibody method, it is characterised in that adopt following steps:
A. the synthesis of feature diagnosing sequence: adopt Fmoc polypeptide solid-state reaction method synthesis feature diagnosing sequence, and increase cysteine residues for linked reaction at the C end of described feature diagnosing sequence;
B. the coupling of feature diagnosing sequence and carrier: select hemocyanin as carrier proteins, connects method with SPDP and described feature diagnosing sequence and described hemocyanin carry out coupling obtains complete antigen;
C. initial immunity: the complete antigen that step b is obtained is diluted with physiological saline, by the complete antigen after dilution and complete Freund's adjuvant 1:(0.8-1.1 by volume) mix, then add penicillin and Streptomycin sulphate carries out emulsification, obtain initial immunity antigen emulsion; Use initial immunity antigen emulsion that rabbit is carried out subcutaneous multi-point injection, carry out initial immunity;
D. antiserum prepare: the complete antigen obtained by step b with physiological saline dilutes, by the complete antigen after dilution and incomplete Freund's adjuvant 1:(0.8-1.1 by volume) mix, then add penicillin and Streptomycin sulphate emulsification, obtain strong immunizing antigen emulsion; After initial immunity the 3rd week and when the 5th week, use strong immunizing antigen emulsion that rabbit is carried out subcutaneous multi-point injection respectively, carry out booster immunization; After carrying out initial immunity the 7th week and when the 9th week, repeat booster immunization, when the antibody titer in rabbit blood sample reaches 1:20000, collect rabbit blood, and be separated obtained antiserum(antisera);
E. the purifying of goathair detection antibody: albumin A filler is mixed with phosphate buffer soln, vacuumizes 20-25min after stirring, obtain the albumin A filler liquid of bubble-free; Albumin A filler liquid is joined in glass column under air bubble free conditions, obtains albumen post; Albumen post is cleaned; Above-mentioned obtained antiserum(antisera) is diluted, joins after dilution and albumen post carries out loading, repeat loading once; Albumen post is cleaned again, carries out wash-out with glycine solution antagonist, after wash-out, clean albumen post, obtained antibody.
2. as claimed in claim 1 a kind of utilize feature diagnosing sequence prepare goathair detection antibody method, it is characterised in that, the feature diagnosing sequence described in step a is the aminoacid sequence PVSCEATICE as shown in sequence table SEQ IDNo.1.
3. a kind of method utilizing feature diagnosing sequence to prepare goathair detection antibody as claimed in claim 1, it is characterised in that, in step c, the concentration of complete antigen after dilution is 0.8-1.2mg/mL.
4. a kind of method utilizing feature diagnosing sequence to prepare goathair detection antibody as claimed in claim 3, it is characterised in that, steps d dilute after the concentration of complete antigen be 0.25-0.75mg/mL.
5. as claimed in claim 1 a kind of utilize feature diagnosing sequence prepare goathair detection antibody method, it is characterised in that, when carrying out initial immunity, booster immunization, the injection volume of each injection point is 100 μ L.
6. as claimed in claim 1 a kind of utilize feature diagnosing sequence prepare goathair detection antibody method, it is characterized in that, in steps d, being separated the method obtaining antiserum(antisera) from blood is: the blood collected is placed in set at room temperature, then solidificating blood is placed in the warm case of 36-38 DEG C and places 25-35min, the refrigerator 11-13h being placed in 4 DEG C again, makes clot fully shrink and obtain the thick antiserum(antisera) that precipitates out completely; Collecting thick antiserum(antisera), in 4 DEG C of environment, when 3000g, centrifugal 14-16min, gets supernatant liquor, is positioned over-80 DEG C of storages for subsequent use.
7. as claimed in claim 1 a kind of utilize feature diagnosing sequence prepare goathair detection antibody method, it is characterized in that, the detailed process of step e is: mixed by the proportioning of 1.5g/ (6-7) mL with phosphate buffer soln by albumin A filler, vacuumize 20-25min after stirring, obtain the albumin A filler liquid of bubble-free; Then the albumin A filler liquid of 6-7 parts by volume is joined in glass column under air bubble free conditions, obtain albumen post; Being cleaned by albumen post with the phosphate buffer soln of 20 parts by volume, cleaning flow velocity is 60mL/h; With phosphate buffer soln, the antiserum(antisera) of 15 parts by volume is diluted to 20 parts by volume, joins after dilution and albumen post carries out loading, repeat loading once; Again being cleaned by albumen post with the phosphate buffer soln of 30 parts by volume, cleaning flow velocity is 60mL/h; With concentration to be 0.2mol/L, pH be 3.0 glycine solution antagonist carry out wash-out; Wash albumen post with phosphate buffer soln after wash-out, obtained antibody, antibody is loaded in centrifuge tube and stand-by in 4 DEG C of preservations.
8. as claimed in claim 7 a kind of utilize feature diagnosing sequence prepare goathair detection antibody method, it is characterised in that, the concentration of phosphate buffer soln used in step e is 0.05mol/L, pH is 7.4.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509107A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic polypeptide |
| CN104447989A (en) * | 2014-12-31 | 2015-03-25 | 浙江理工大学 | Silkworm fibroin antibody preparing method |
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- 2016-02-24 CN CN201610100033.2A patent/CN105669861A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509107A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic polypeptide |
| CN104447989A (en) * | 2014-12-31 | 2015-03-25 | 浙江理工大学 | Silkworm fibroin antibody preparing method |
Non-Patent Citations (1)
| Title |
|---|
| 周玉枝等: "《惊奇化学》", 31 August 2013, 上海辞书出版社 * |
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Application publication date: 20160615 |