CN105504056B - The method for preparing tussah silk fibroin antibody using feature hexapeptide - Google Patents
The method for preparing tussah silk fibroin antibody using feature hexapeptide Download PDFInfo
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Abstract
The present invention discloses a kind of method for preparing tussah silk fibroin antibody using feature hexapeptide, and composition sequence is the hexapeptide of " GSGAGG ", and hexapeptide and keyhole limpet hemocyanin coupling are got up to obtain comlete antigen；With normal saline dilution comlete antigen, the comlete antigen after dilution is uniformly mixed with Quick Antibody-Rabbit 5W adjuvant, initial immunity then is carried out to rabbit, booster immunization then is carried out to rabbit, the reagent that booster immunization uses is consistent with initial immunity；When the potency of the antibody in rabbit blood sample reaches 1:10000, collect it is immune after rabbit blood, and make that clot is sufficiently shunk and antiserum is precipitated completely, then collect antiserum and centrifugal treating obtains supernatant, packing is spare.The method of the present invention is simple and fast, and the antibody prepared has very strong specificity, can be used to detect the fibroin albumen in textile.
Description
Technical field
The present invention relates to the preparation methods of polyclonal antibody more particularly to a kind of utilization feature polypeptide to prepare tussah silk peptide egg
The method of Bai Kangti.
Background technique
Silk is a kind of natural polymer protein fibre, is broadly divided into domestic silkworm silk and tussah two major classes.Wild silkworm also cries
Tussah is gained the name because being fond of mongolian oak leaf, cocoon can filature, be mainly used for weave tussah silk.China is to utilize tussah earliest and put toothed oak in a suitable place to breed
The country of silkworm, but its origin has been a hot spot of research difficult point, and there is an urgent need to a kind of more scientific methods to identify early stage grave
Or the silk goods in traces.The method of previous identification Ancient Silk Textile is also relatively more, but silk is as a kind of organic polymer material
Material is chronically in grave or ruins environment, will necessarily be influenced by factors, be degraded to silk protein occur
And the problems such as macromolecular chain break, it is difficult to detect the presence of fibroin albumen using traditional certain methods.Therefore how to adopt
With the advanced means of natural science, silk goods microscratch detection technique system is established, from mark, residue extracts ancient times in soil
The information of silk, it is very urgent to the research of silk origin.The advanced technology of protein detection is based on antibody-antigene at present
Western Blotting or ELISA method, the preparation of tussah silk fibroin antibody are a key links of the research.
Summary of the invention
The present invention is in order to overcome the above-mentioned deficiencies of the prior art, and it is an object of the present invention to provide a kind of prepare toothed oak using feature polypeptide
The method of fibroin protein antibody, it is simple and quick, and the antibody prepared has very strong specificity.
To achieve the above object, the technical solution that the present invention uses is: a kind of to prepare tussah silk using feature hexapeptide
The method of fibroin antibody, it is characterized in that, the following steps are included:
Step 1: it is the polypeptide of " CGSGAGG " using Fmoc method composition sequence, and passes through half Guang of the N-terminal of told polypeptide
Propylhomoserin makes the polypeptide and keyhole limpet hemocyanin be coupled to obtain comlete antigen；
Step 2: the comlete antigen described in normal saline dilution, by the comlete antigen and Quick Antibody- after dilution
Rabbit 5W adjuvant (Quick Antibody series immunologic adjuvant, commercially available on the market) uniformly mixes, and obtains used in initial immunity
Reagent；
It chooses 2 White Rabbits and carries out initial immunity and booster immunization, extract rabbit ear blood sample before immune and compare；
Initial immunity is carried out to rabbit first, booster immunization then carried out to rabbit, reagent that booster immunization uses and just
It is secondary immune consistent；When the potency of the antibody in rabbit blood sample reaches 1:10000, the rabbit blood after being immunized is collected, and make blood
Block is sufficiently shunk and antiserum is precipitated completely, then collects antiserum and centrifugal treating obtains supernatant, dispense spare；
Step 3: it is purified using immunoaffinity chromatography technical antagonism serum.First by haptens (the i.e. polypeptide of synthesis
" GSGAGG ") and chromatographic stuffing crosslinking aid S ulfo-link-gel (Pierce company, the U.S. production, commercially available on the market) handed over
Connection, closes the glycine of " GSGAGG " N-terminal；
Column is pre-processed with the PBS of 20-25ml, 50mM, pH7.4, pretreated flow velocity is 60-65ml/h.
The antiserum of 15ml is diluted to 20ml, loading after dilution with the PBS of 50mM, pH7.4.It is primary to repeat loading.
Column is cleaned with the PBS of 30-35ml, 50mM, pH7.4, the flow velocity of cleaning is 60-65ml/h.With pH3.0,
The glycine-HCL (glycine hydrochloride) of 0.1M elutes antibody.It used after the completion of elution containing 50mM, pH7.4, contain 0.02%
The PBS of merthiolate washs polypeptide column, and the antibody obtained after purification adjusts pH to 7.4 and stores in 3-5 DEG C；
Step 4: antibody specificity effect measuring: antigen fibroin albumen is coated with the concentration of 1 μ g/ml, and 4 DEG C of coatings are overnight.
Antibody is diluted to 240000 times, 60000 times, 20000 times, 10000 times, 5000 times, 1000 times, 200 times respectively, 50 holes μ l/
Loading is measured, incubates 1h at 37-39 DEG C.With TBST (by 8 grams of NaCl, KCl 0.2g, 3 grams of Trise base, add pure water 800ML,
With concentrated hydrochloric acid tune PH to 7.4, then plus water is to 1000ML, obtains TBS solution；In TBS plus 0.1% polysorbas20, i.e., 0.1% washes
Wash liquid) with 200 holes μ l/ washing 1-3 times.ELIAS secondary antibody is diluted with 1:5000 to be used, and 50 holes μ l/ incubate 1h at 37-39 DEG C.With
TBST is with 200 holes μ l/ washing 2-4 times.TMB is added with 100 holes μ l/ to develop the color in shady place, after the 10min that develops the color, with 100 holes μ l/
The H of 2M is added2SO4It terminates；
Step 5: OD is measured with microplate reader450Value；
Work as OD450It is positive then to prove that sample detected is presented, illustrates to contain fibroin albumen in sample by value > 0.6；
Work as OD450It is negative then to prove that measured sample is presented, illustrates in sample without containing fibroin albumen for value≤0.6.
In the step 2, comlete antigen is diluted to 2 times of ultimate densities with physiological saline.
In the step 2, prepare the comlete antigen for the first time and when booster immunization reagent, after dilution and adjuvant be by
Volume ratio 1:1 is mixed.
In the step 2, the method for carrying out initial immunity to rabbit is: by back leg Calf muscle injecting immune rabbit
Son, every rabbit inject 100 μ l；The method for carrying out booster immunization to rabbit is: at the 21st day of initial immunity by the same manner
One needle of booster immunization.
In the step 2, collect antiserum post-processing method be: temperature be 4 DEG C in 3000g under the conditions of be centrifuged
15min dispenses supernatant, is placed in -80 DEG C and stores for future use.
Compared with prior art, the invention has the advantages that:
(1) the method for the present invention has very strong special by polyclonal antibody prepared by the feature polypeptide of tussah silk fibroin
Property, through enzyme-linked immunization analysis it is found that the antibody can have apparent immune response with tussah silk fibroin, sensitivity is very
Height can be used for detecting tussah silk fibroin, the detection and analysis particularly suitable for imperfect fibroin albumen.
(2) different from the antibody that conventional method is prepared using the macromolecular of native protein, antibody prepared by the present invention is not
It is only able to detect complete fibroin albumen, can be used for the silk fibroin molecular being broken in detection such as Ancient Silk Textile,
A kind of sensitive special efficiently recognition methods is provided to remain product for the silk weaving in grave of Chinese early stage and traces, is examined for silk
Ancient and Origin provides new scientific basis.
(3) immunologic adjuvant that the present invention uses, it is nontoxic, protein component is free of, does not destroy antigen native configurations, and
And immunization ways are simple, without emulsifying antigen (only needing to be simply mixed and can be immunized), while the immune period is short, antigen dosage
It is few.
Specific embodiment
It elaborates below to the embodiment of the present invention.Following embodiment is only further described the application, no
It is interpreted as the limitation to the application.
The amino acid of tussah silk fibroin is checked on NCBI (http://www.ncbi.nlm.nih.gov/pubmed/)
Sequence.
By statistical analysis, determines that " GSGAGG " polypeptide fragment frequency of occurrences is higher, be the feature polypeptide of tussah silk peptide, therefore
Antigen is prepared with this segment.
Detailed description below preparation method of the invention.
Embodiment 1:
A method of preparing tussah silk fibroin antibody using feature hexapeptide, it the following steps are included:
Comlete antigen synthesis:
Haptens (i.e. " CGSGAGG " polypeptide sequence) is synthesized using Fmoc method, is added in the N-terminal of " CGSGAGG " polypeptide sequence
Cysteine is added to be coupled the polypeptide and keyhole limpet hemocyanin (KLH).Pass through the effect of crosslinking agent, half Guang of " CGSGAGG " N-terminal
Sulfydryl and KLH primary amine on propylhomoserin form covalent bond, so that polypeptide and KLH coupling be got up, obtain comlete antigen.
Comlete antigen is diluted to 2 times of ultimate densities with physiological saline, by the comlete antigen and Quick after dilution
1:1 is uniformly mixed Antibody-Rabbit 5W adjuvant by volume, obtains reagent used in initial immunity.Booster immunization uses
Reagent it is consistent with initial immunity.
Antibody preparation:
It chooses 2 White Rabbits (15 week old or so) and carries out initial immunity and booster immunization.Rabbit ear blood sample is first extracted before immune
It compares.Initial immunity is by back leg Calf muscle injecting immune rabbit, and every rabbit injects 100 μ l.Then exempt from for the first time
One needle of the same manner booster immunization is pressed with same reagent in the 21st day of epidemic disease.The 10th day venous blood sampling inspection after being immunized at second
Survey antiserum titre.
When the sero-fast potency in rabbit blood sample reaches 1:10000, kills rabbit and collect blood, a large amount of blood of collection
Liquid is set allows it to solidify at room temperature；Solidificating blood is placed in 37 DEG C of incubators and places 30min, then is placed in 4 DEG C of refrigerator overnights, makes clot
It sufficiently shrinks and antiserum is precipitated completely.Antiserum is collected, 3000g is centrifuged 10min at 4 DEG C, dispenses supernatant, puts -80 DEG C of storages
It deposits spare.
Antibody purification:
It is purified using immunoaffinity chromatography technical antagonism serum.First by the haptens of synthesis (i.e. polypeptide " GSGAGG ")
It is crosslinked with chromatographic stuffing Sulfo-link-gel, closes the glycine of " GSGAGG " N-terminal.With 20ml, 50mM, pH7.4
PBS column is pre-processed, pretreated flow velocity be 60ml/h.The antiserum of 15ml is diluted with the PBS of 50mM, pH7.4
To 20ml, loading after dilution.It is primary to repeat loading.Column is cleaned with the PBS of 30ml, 50mM, pH7.4, the flow velocity of cleaning
For 60ml/h.Antibody is eluted with the glycine-HCL of pH3.0,0.1M.Elution after the completion of with contain 50mM, pH7.4, contain
The PBS of 0.02% merthiolate washs polypeptide column, and the antibody obtained after purification adjusts pH to 7.4 and stores in 3 DEG C.
Antibody specificity effect measuring: antigen fibroin albumen is coated with the concentration of 1 μ g/ml, and 4 DEG C of coatings are overnight.By antibody
240000 times, 60000 times, 20000 times, 10000 times, 5000 times, 1000 times, 200 times of dilution respectively, the amount loading in 50 holes μ l/,
1h is incubated at 37 DEG C.It is washed 2 times with TBST with 200 holes μ l/.ELIAS secondary antibody is diluted with 1:5000 to be used, 50 holes μ l/, at 37 DEG C
Incubate 1h.It is washed 3 times with TBST with 200 holes μ l/.TMB is added with 100 holes μ l/ to develop the color in shady place, after the 10min that develops the color, with
The H of 100 holes μ l/ addition 2M2SO4It terminates.OD is measured with microplate reader450Value.According to OD450Value, situations such as to the combination of antigen-antibody
Judged.Work as OD450Value is assured that in sample when reaching positive criteria contains fibroin；If being free of fibroin in sample,
OD450Value will be lower than positive criteria, can quickly judge the presence of fibroin accordingly.
Interpretation of result: as the result is shown in addition to blank control is in colourless, sample well is in yellow.OD is measured with microplate reader450Value
It as a result is that 240000 times, 60000 times, 20000 times, 10000 times, 5000 times, 1000 times, 200 times are carried out to antibody after purification
The OD obtained after dilution450Value is respectively 0.370,0.700,1.092,1.491,1.606,2.109,2.145, blank control
OD450Value is 0.078. according to platform OD450Value reaches 0.6 for positive requirement, even if by 60000 times of the antibody dilution of preparation,
It still is able to detect the positive signal of fibroin albumen.It can illustrate from result above, by the antibody of the method for the present invention preparation to silk
The reaction sensitivity of element is high, and easy to operate.
Embodiment 2:
A method of preparing tussah silk fibroin antibody using feature hexapeptide, it the following steps are included:
Comlete antigen synthesis:
Haptens (i.e. " CGSGAGG " polypeptide sequence) is synthesized using Fmoc method, is added in the N-terminal of " CGSGAGG " polypeptide sequence
Cysteine is added to be coupled the polypeptide and keyhole limpet hemocyanin (KLH).Pass through the effect of crosslinking agent, half Guang of " CGSGAGG " N-terminal
Sulfydryl and KLH primary amine on propylhomoserin form covalent bond, so that polypeptide and KLH coupling be got up, obtain comlete antigen.
Comlete antigen is diluted to 2 times of ultimate densities with physiological saline, by the comlete antigen and Quick after dilution
1:1 is uniformly mixed Antibody-Rabbit 5W adjuvant by volume, obtains reagent used in initial immunity.Booster immunization uses
Reagent it is consistent with initial immunity.
Antibody preparation:
It chooses 2 White Rabbits (15 week old or so) and carries out initial immunity and booster immunization.Rabbit ear blood sample is first extracted before immune
It compares.Initial immunity is by back leg Calf muscle injecting immune rabbit, and every rabbit injects 100 μ l.Then exempt from for the first time
One needle of the same manner booster immunization is pressed with same reagent in the 21st day of epidemic disease.The 10th day venous blood sampling inspection after being immunized at second
Survey antiserum titre.
When the sero-fast potency in rabbit blood sample reaches 1:10000, kills rabbit and collect blood, a large amount of blood of collection
Liquid is set allows it to solidify at room temperature；Solidificating blood is placed in 37 DEG C of incubators and places 30min, then is placed in 4 DEG C of refrigerator overnights, makes clot
It sufficiently shrinks and antiserum is precipitated completely.Antiserum is collected, 3000g is centrifuged 10min at 4 DEG C, dispenses supernatant, puts -80 DEG C of storages
It deposits spare.
Antibody purification:
It is purified using immunoaffinity chromatography technical antagonism serum.First by the haptens of synthesis (i.e. polypeptide " GSGAGG ")
It is crosslinked with chromatographic stuffing Sulfo-link-gel, closes the glycine of " GSGAGG " N-terminal.
Column is pre-processed with the PBS of 25ml, 50mM, pH7.4, pretreated flow velocity is 65ml/h.With 50mM,
The antiserum of 15ml is diluted to 20ml, loading after dilution by the PBS of pH7.4.It is primary to repeat loading.
Column is cleaned with the PBS of 35ml, 50mM, pH7.4, the flow velocity of cleaning is 65ml/h.With pH3.0,0.1M
Glycine-HCL elutes antibody.It is washed after the completion of elution with containing 50mM, pH7.4, the PBS containing 0.02% merthiolate
Polypeptide column, the antibody obtained after purification adjust pH to 7.4 and store in 5 DEG C.
Antibody specificity effect measuring: antigen fibroin albumen is coated with the concentration of 1 μ g/ml, and 4 DEG C of coatings are overnight.By antibody
240000 times, 60000 times, 20000 times, 10000 times, 5000 times, 1000 times, 200 times of dilution respectively, the amount loading in 50 holes μ l/,
1h is incubated at 39 DEG C.It is washed 3 times with TBST with 200 holes μ l/.ELIAS secondary antibody is diluted with 1:5000 to be used, 50 holes μ l/, at 39 DEG C
Incubate 1h.It is washed 4 times with TBST with 200 holes μ l/.TMB is added with 100 holes μ l/ to develop the color in shady place, after the 10min that develops the color, with
The H of 100 holes μ l/ addition 2M2SO4It terminates.OD is measured with microplate reader450Value.According to OD450Value, situations such as to the combination of antigen-antibody
Judged.Work as OD450Value is assured that in sample when reaching positive criteria contains fibroin；If being free of fibroin in sample,
OD450Value will be lower than positive criteria, can quickly judge the presence of fibroin accordingly.
Interpretation of result: as the result is shown in addition to blank control is in colourless, sample well is in yellow.OD is measured with microplate reader450Value
It as a result is that 240000 times, 60000 times, 20000 times, 10000 times, 5000 times, 1000 times, 200 times are carried out to antibody after purification
The OD obtained after dilution450Value is respectively 0.370,0.700,1.092,1.491,1.606,2.109,2.145, blank control
OD450Value is 0.078. according to platform OD450Value reaches 0.6 for positive requirement, even if by 60000 times of the antibody dilution of preparation,
It still is able to detect the positive signal of fibroin albumen.It can illustrate from result above, by the antibody of the method for the present invention preparation to silk
The reaction sensitivity of element is high, and easy to operate.
Claims (3)
1. a kind of method for preparing tussah silk fibroin antibody using feature hexapeptide, it is characterized in that, the following steps are included:
Step 1: it is the polypeptide of " CGSGAGG " using Fmoc method composition sequence, and passes through the cysteine of the N-terminal of the polypeptide
The polypeptide and keyhole limpet hemocyanin is set to be coupled to obtain comlete antigen；
Step 2: the comlete antigen described in normal saline dilution, by the comlete antigen and Quick Antibody- after dilution
Rabbit 5W adjuvant uniformly mixes, and obtains reagent used in initial immunity；Comlete antigen is diluted to 2 times most with physiological saline
Final concentration；When preparing first and booster immunization reagent, comlete antigen after dilution is that 1:1 is mixed by volume with adjuvant
It closes, chooses 2 White Rabbits and carry out initial immunity and booster immunization, extract rabbit ear blood sample before immune and compare；
First to rabbit carry out initial immunity, then to rabbit carry out booster immunization, the reagent that booster immunization uses with exempt from for the first time
Epidemic disease is consistent；When the potency of the antibody in rabbit blood sample reaches 1:10000, the rabbit blood after being immunized is collected, and fill clot
Divide contraction and antiserum to be precipitated completely, then collects antiserum and centrifugal treating obtains supernatant, dispense spare；
Step 3: it is purified using immunoaffinity chromatography technical antagonism serum；First by haptens (the i.e. polypeptide of synthesis
" GSGAGG ") and chromatographic stuffing crosslinking aid S ulfo-link-gel be crosslinked, close the glycine of " GSGAGG " N-terminal；
Column is pre-processed with the PBS of 20ml, 50mM, pH7.4, pretreated flow velocity is 60ml/h；
The antiserum of 15ml is diluted to 20ml with the PBS of 50mM, pH7.4, it is primary to repeat loading for loading after dilution；
Column is cleaned with the PBS of 30ml, 50mM, pH7.4, the flow velocity of cleaning is 60ml/h；With the sweet ammonia of pH3.0,0.1M
Hydrochlorate acid glycine-HCL elutes antibody；After the completion of elution with containing 50mM, pH7.4, containing 0.02% merthiolate
PBS column scrubber, the antibody obtained after purification adjust pH to 7.4 and store in 4 DEG C；
Step 4: antibody specificity effect measuring: antigen fibroin albumen is coated with the concentration of 1 μ g/ml, and 4 DEG C of coatings are overnight；It will resist
Body dilutes 240000 times, 60000 times, 20000 times, 10000 times, 5000 times, 1000 times, 200 times respectively, in the amount in 50 holes μ l/
Sample incubates 1h at 37 DEG C；It is washed 2 times with TBST with 200 holes μ l/；ELIAS secondary antibody is diluted with 1:5000 to be used, 50 holes μ l/, 37 DEG C
Lower incubation 1h；It is washed 3 times with TBST with 200 holes μ l/；TMB is added with 100 holes μ l/ to develop the color in shady place, after the 10min that develops the color, with
The H2SO4 that 2M is added in 100 holes μ l/ is terminated；
Step 5: 450 value of OD is measured with microplate reader；
Work as OD450It is positive then to prove that sample detected is presented, illustrates to contain fibroin albumen in sample by value > 0.6；Work as OD450Value
≤ 0.6, then it is negative to prove that measured sample is presented, illustrates in sample without containing fibroin albumen.
2. according to the method described in claim 1, it is characterized in that: in the step 2, the side of initial immunity is carried out to rabbit
Method is: by back leg Calf muscle injecting immune rabbit, every rabbit injects 100 μ l；The method that booster immunization is carried out to rabbit
It is: at the 21st day of initial immunity by one needle of the same manner booster immunization.
3. according to the method described in claim 1, it is characterized in that: in the step 2, collecting the method that antiserum post-processes
Be: temperature be 4 DEG C in 3000g under the conditions of be centrifuged 15min, dispense supernatant, be placed in -80 DEG C and store for future use.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509108A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic dodecapeptide |
| 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 |
| CN104232665A (en) * | 2014-07-23 | 2014-12-24 | 湖州南浔恒岳农业科技有限公司 | Silk protein peptide fragment for identifying novel natural color silkworm and preparation method of silk protein peptide fragment |
| CN104447989A (en) * | 2014-12-31 | 2015-03-25 | 浙江理工大学 | Silkworm fibroin antibody preparing method |
-
2015
- 2015-11-12 CN CN201510772674.8A patent/CN105504056B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103509108A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic dodecapeptide |
| 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 |
| CN104232665A (en) * | 2014-07-23 | 2014-12-24 | 湖州南浔恒岳农业科技有限公司 | Silk protein peptide fragment for identifying novel natural color silkworm and preparation method of silk protein peptide fragment |
| CN104447989A (en) * | 2014-12-31 | 2015-03-25 | 浙江理工大学 | Silkworm fibroin antibody preparing method |
Non-Patent Citations (2)
| Title |
|---|
| 丝素蛋白亚基研究;于同隐等;《高分子学报》;19971031(第5期);629-632 * |
| 丝素蛋白的构象与结晶性;闻荻江等;《纺织学报》;20050228;第26卷(第1期);110-112 * |
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Inventor after: Wang Bing Inventor after: Li Jin Inventor after: You Qiushi Inventor after: Peng Zhiqin Inventor after: Hu Zhiwen Inventor before: You Qiushi Inventor before: Wang Bing Inventor before: Peng Zhiqin Inventor before: Hu Zhiwen |
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