Gamma S-crystalin protein and preparation method of egg yolk antibody of gamma S-crystalin protein

By expressing and purifying γS-crystallin protein antigen in E. coli and extracting antibodies in combination with PEG6000 precipitation method, the shortcomings in the preparation of γS-crystallin protein antigen in the prior art were solved, and efficient and specific antibody preparation and detection were achieved, promoting the promotion of Early diagnosis and prevention of cataract disease.

CN119285740APending Publication Date: 2025-01-10SHAANXI SCI TECH UNIV
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Patent Information

Application Number
CN202411503835.9
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Priority Date
2024-10-14
Filing Date
2024-10-25
Publication Date
2025-01-10

AI Technical Summary

Technical Problem

The prior art is difficult to effectively prepare γS-crystallin protein antigen and its yolk antibodies, which limits the early diagnosis and treatment of diseases such as cataracts.

Method used

By downloading the γS-crystallin protein sequence from NCBI, recombinant plasmids were constructed and purified antigens were expressed in E. coli, avian polyclonal antibody IgY was extracted in combination with PEG6000 precipitation, and an indirect competitive enzyme-linked immunoassay was established.

Benefits of technology

It provides efficient, specific and affinity γS-crystallin protein antibodies for early diagnosis and prevention of cataract diseases, improves the rapid, sensitive and accurate detection, and provides a theoretical basis for future disease treatment and prevention.

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Abstract

The invention discloses a preparation method of a gamma S-crystalin protein antigen and a preparation method of an egg yolk antibody of the gamma S-crystalin protein antigen. The preparation method comprises the following steps: preparing the gamma S-crystalin protein antigen by adopting a gene recombination method, and emulsifying the gamma S-crystalin protein antigen; healthy laying hens are selected for injection immunization; an egg yolk antibody IgY is extracted from an egg obtained from an immunized laying hen, and an enzyme-linked immunosorbent assay method for detecting gamma S-crystalin in a sample is established at the same time. An expression plasmid containing gamma S-crystalin protein is constructed and expressed in BL21 (DE3) escherichia coli, an expression product obtained after separation and purification serves as an antigen and is mixed with a Freund's adjuvant to be emulsified, then laying hens are immunized, eggs are collected, the egg yolk antibody IgY in egg yolk is extracted, and the antibody is natural immune globulin. According to the invention, a polyclonal antibody technology is combined with an ELISA method, and the prepared anti-egg yolk antibody IgY can be used for efficiently detecting the existence of gamma S-crystalin protein. The method is suitable for preparing the gamma S-crystalin egg yolk antibody IgY, and the obtained antibody is safe and high in titer and has good development and application prospects.
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