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Oriental schistosomiasis resistant natural numerator vaccine specific single-chain antibody

A single-chain antibody and specific technology, applied in the field of specific anti-schistosomiasis single-chain antibody, can solve the problems of reducing therapeutic effect, causing human anti-mouse reaction, causing allergic reaction or hypersensitivity reaction, etc.

Inactive Publication Date: 2009-05-13
何卓 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, making monoclonal antibodies requires animals, specialized cell culture equipment and a lot of time and effort
When applied to human treatment, as a foreign protein, mouse-derived monoclonal antibodies can trigger human anti-mouse reactions, thereby reducing or eliminating the therapeutic effect, and even triggering allergic reactions or hypersensitivity reactions

Method used

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  • Oriental schistosomiasis resistant natural numerator vaccine specific single-chain antibody
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  • Oriental schistosomiasis resistant natural numerator vaccine specific single-chain antibody

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Embodiment Construction

[0011] 1. Animal immunization and total RNA extraction

[0012] SIEA was thoroughly mixed with an equal volume of Freund's complete adjuvant, and 100ug of SIEA was injected intraperitoneally into each mouse. After two weeks, switch to Freund's incomplete adjuvant and mix well with equal volume of SIEA, strengthen once every two weeks, a total of four times. On the seventh day after the first immunization, the tail vein blood of the mice was taken to measure the titer by ELISA. The mouse spleen with a titer of more than 10,000 was taken, and the total RNA of the spleen was extracted with Trizol reagent.

[0013] 2. Synthesis of the first strand of cDNA and amplification of a complete set of VH and VL gene fragments

[0014] According to the instructions of the RT kit from MBI Company, mouse spleen RNA was used as a template, oligo(dt)18 was used as a primer, and the first strand of cDNA was synthesized by reverse transcription with M-mulv reverse transcriptase. Then using cD...

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Abstract

The invention relates to the field of genetic engineering antibody, and builds schistosoma japonicum immature egg soluble antigen (SIEA) single-chain antibody library for the first time. By screening the SIEA single-chain antibody library through an immuno-affinity screening technique, a specific single-chain antibody aiming at anti-schistosomiasis-japonica effective natural molecular vaccine SIEA26-28kDa is obtained. The obtainment of the specific single-chain antibody provides a foundation for treating schistosomiasis japonica, utilizing the specific single-chain antibody as a probe to screen schistosoma japonicum cDNA libraries and obtaining a corresponding gene sequence aiming at anti-schistosomiasis-japonica effective vaccine molecules more conveniently and rapidly.

Description

Technical field: [0001] The present invention relates to the field of genetically engineered antibodies, in particular to a specific anti-schistosomiasis single-chain antibody, which can be directly used as a passive animal test with or without modification (adding immunotoxin or anti-schistosomiasis drug) , to observe the treatment effect of schistosomiasis; this single-chain antibody can also be used to screen the cDNA library of Schistosoma japonicum to obtain the nucleotide sequence encoding antigen molecule SIEA26-28kDa, which provides the possibility for mass production of anti-Schistosomiasis vaccine. Background technique: [0002] Schistosomiasis is still a zoonotic disease that seriously endangers human health and life safety and hinders economic development and social progress in infected areas. Chitsulo and others reported that 600 million people in 74 countries around the world were threatened with infection, 200 million people were infected, 120 million people s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/06C12N15/13C40B40/10
Inventor 何卓汪世平曾少华刘明社徐绍锐
Owner 何卓
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