A kind of preparation method of silk fibroin antibody of silkworm

A silk fibroin and silk technology, applied in the direction of serum immunoglobulin, anti-animal/human immunoglobulin, etc., can solve problems such as difficult to detect the existence of silk fibroin, difficult to explore the origin of silk, and hard to find evidence , to achieve the effect of high sensitivity, strong specificity and fast identification method

Active Publication Date: 2017-12-12
ZHEJIANG SCI-TECH UNIV
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Problems solved by technology

In the past, there were many methods for identifying ancient silk fabrics. However, due to the influence of various external factors, such as light, acid, alkali, microorganisms and other environmental factors, most of the silk macromolecules have been degraded into peptides and amino acids. It is difficult to detect the presence of silk fibroin, so it is difficult to trace the origin of silk, and the older the evidence is, the harder it is to find

Method used

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Embodiment Construction

[0014] Take the following steps:

[0015] A) Extract silk fibroin: the mulberry silk is mixed with NaCO with a mass concentration of 0.5% at a bath ratio of 1:100 3 Boil in the solution for 30 minutes for degumming, and obtain silk fibroin after degumming twice; wash the silk fibroin with deionized water, and place it in an oven at 40-60°C for 12 hours; dry the silk fibroin at 98±2°C In the water bath, CaCl with a mass concentration of 50% was used at a bath ratio of 1:25 2 The solution was dissolved for 1.5h; the CaCl in which the silk fibroin was dissolved 2 The solution was naturally cooled to room temperature, then filtered, dialyzed with a dialysis bag with a molecular weight cut-off of 8000-14000, and deionized water was changed every 4 hours, and a pure silk fibroin solution was obtained after 72 hours; the silk fibroin solution was concentrated and then freeze-dried 24h; Obtain the silk fibroin that sticks into a block, and obtain silk fibroin powder after grinding; ...

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Abstract

The invention discloses a method for preparing silkworm silk fibroin antibody. The calcium chloride ethanol system is used to extract the silk fibroin, and then the silk fibroin is injected into rabbits as a complete antigen for animal immunization to obtain the silk fibroin antibody. The steps are as follows: silk fibroin extraction: extraction with calcium chloride ethanol system; peptide immunization and antiserum preparation: mixing and emulsifying silk fibroin powder with Freund's adjuvant, injecting multiple points subcutaneously in rabbits, and repeatedly boosting immunization until Take antiserum to detect that the antibody titer reaches the standard and stop immunization; antibody purification: after the antibody titer of the rabbit reaches the standard, take blood to make the blood clot fully coagulate, and separate the antiserum; the effectiveness of the silk fibroin antibody is determined by the ELISA step For identification, the obtained silk fibroin antibody is used as the primary antibody, and the standard enzyme-linked immunosorbent method is used to confirm that the silk fibroin antibody can interact with silk fibroin, which can be used in the enzyme-linked immunosorbent assay.

Description

technical field [0001] The invention belongs to the field of biological antibodies, in particular to a method for preparing silk fibroin polyclonal antibodies by using silk fibroin. Background technique [0002] The silkworm is a silk-spinning insect with high economic value, mainly composed of silk fibroin and sericin. However, sericin is easy to fall off during the silk reeling process, so the identification of silk is mainly the identification of silk fibroin. As the crystallization of human wisdom, the research on the origin of silk has become more and more popular in recent years. There is an urgent need for a more scientific method to identify silk fabrics in early tombs. In the past, there were many methods for identifying ancient silk fabrics. However, due to the influence of various external factors, such as light, acid, alkali, microorganisms and other environmental factors, most of the silk macromolecules have been degraded into peptides and amino acids. The pre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C07K16/06
Inventor 王秉刘苗苗
Owner ZHEJIANG SCI-TECH UNIV
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