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Method for using characteristic hexapeptide for preparing tussah fibroin protein antibody

A technology of tussah silk fibroin and antibody, which is applied in the preparation method of peptides, chemical instruments and methods, and from serum immunoglobulin, etc. problems, to achieve the effect of short immunization cycle, high sensitivity and less antigen dosage

Active Publication Date: 2016-04-20
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows creating highly specialized proteins called chitraxidase or talanum spermine oxazolidone polymeric microspheres which are able to attach specifically to certain types of silks found on different parts of human body like skin, hair, nails, teeth etc., making them easier to see even when they're worn down over time due to their unique properties. These tiny particles help identify damaged areas caused during manufacturing processes. They may include other substances added into cosmetic products meant to make it look better than regular ones.

Problems solved by technology

This patented technical problem addressed in this patents relates to identifying old silks that have become lost during storage due to environmental influences such as weather conditions (temperature) and other causes. Current techniques involve analyzers with expensive equipment, which makes them hard to operate at remote locations where archaeology remains hidden behind fossil records containing these materials.

Method used

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  • Method for using characteristic hexapeptide for preparing tussah fibroin protein antibody
  • Method for using characteristic hexapeptide for preparing tussah fibroin protein antibody

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Embodiment 1

[0030] A method for preparing tussah silk fibroin antibody using characteristic hexapeptides, which includes the following steps:

[0031] Complete antigen synthesis:

[0032] The hapten (ie, "CGSGAGG" polypeptide sequence) is synthesized by the Fmoc method, and cysteine ​​is added to the N end of the "CGSGAGG" polypeptide sequence to couple the polypeptide with keyhole limpet hemocyanin (KLH). Through the action of the cross-linking agent, the sulfhydryl group on the N-terminal cysteine ​​of "CGSGAGG" and the primary KLH amine form a covalent bond, thereby coupling the polypeptide and KLH to obtain a complete antigen.

[0033] Dilute the complete antigen to 2 times the final concentration with normal saline, and mix the diluted complete antigen with QuickAntibody-Rabbit5W adjuvant in a volume ratio of 1:1 to obtain the reagents for the first immunization. The reagents used in the booster immunization are the same as the initial immunization.

[0034] Antibody preparation:

[0035] Tw...

Embodiment 2

[0042] A method for preparing tussah silk fibroin antibody using characteristic hexapeptides, which includes the following steps:

[0043] Complete antigen synthesis:

[0044] The hapten (ie, "CGSGAGG" polypeptide sequence) is synthesized by the Fmoc method, and cysteine ​​is added to the N end of the "CGSGAGG" polypeptide sequence to couple the polypeptide with keyhole limpet hemocyanin (KLH). Through the action of the cross-linking agent, the sulfhydryl group on the N-terminal cysteine ​​of "CGSGAGG" and the primary KLH amine form a covalent bond, thereby coupling the polypeptide and KLH to obtain a complete antigen.

[0045] Dilute the complete antigen to 2 times the final concentration with normal saline, and mix the diluted complete antigen with QuickAntibody-Rabbit5W adjuvant in a volume ratio of 1:1 to obtain the reagents for the first immunization. The reagents used in the booster immunization are the same as the initial immunization.

[0046] Antibody preparation:

[0047] Tw...

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Abstract

The present invention discloses a method for using a characteristic hexapeptide for preparing a tussah fibroin protein antibody, a synthetic sequence is a ''GSGAGG'' hexapeptide, the hexapeptide is coupled with keyhole limpet hemocyanin to obtain a full antigen; the full antigen is diluted with normal saline, the diluted full antigen is mixed evenly with Quick Antibody-Rabbit 5 w adjuvant, primary immunization is performed on a rabbit, then booster immunization is performed on the rabbit, reagents used for the booster immunization and the primary immunization are same; when titer of the antibody in a blood sample of the rabbit reaches 1: 10,000, blood of the immunized rabbit is collected, blood clots are fully contracted, antiserum is completely precipitated, then the antiserum is collected antiserum and centrifuged to obtain a supernatant, and the supernatant is sub-packed for standby use. The method is simple and quick, and the prepared antibody is highly specific and can be used to detect silk fibroin in textiles.

Description

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Claims

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Application Information

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Owner ZHEJIANG SCI-TECH UNIV
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