Anti-schistosoma japonicum monoclonal antibody NP11-4 chimeric Fab antibody, preparation method and use

Abstract

The invention discloses a monoclonal antibody NP11-4 chimeric Fab antibody against schistosoma japonicum, a preparation method and the application thereof, which relates to genetic engineering technology and the field of preparing therapeutic drugs for schistosomiasis. The application is located on the monoclonal antibody NP11-4 of surface membrane of adult worms, cercaria membrane, schistosomulum membrane, egg shell and miracidium membrane of the schistosoma japonicum; genetic engineering technology is used for extracting the total RNA of a hybrid tumor cell strain; an RT-PCR method is used for amplifying VH and VL genes; PCR is used for respectively amplifying the Fd and the total length L of murine VH, VL and human CH1of IgG1, CL, and then the chimeric Fab gene is amplified by overlapping and extending PCR and using Fd and L genes as templates; the chimeric Fab gene is restricted enzyme and connected with a carrier pComb3XSS and cloned to colon bacillus Top10F', and then an interest protein is induced to acquire soluble expression. At the N end of the expressed monoclonal antibody NP11-4 chimeric Fab antibody against schistosoma japonicum, the part from the first amino acid to the 215th amino acid is the L chain of the chimeric Fab antibody, and the part from the 216th amino acid to the 488th amino acid is the Fd section of the chimeric Fab antibody, the L chain and the Fd section are connected through a disulfide linkage, and the chimeric Fab antibody has an overall length of 488 amino acids.

Description

technical field [0001] The invention relates to an anti-schistosomiasis monoclonal antibody NP11-4 chimeric Fab antibody and its preparation method and application, and uses genetic engineering technology to prepare the anti-schistosomiasis chimeric Fab antibody. The chimeric Fab variable region gene fragment comes from the monoclonal antibody NP11-4 located on the surface membrane, cercariae membrane, juvenile membrane, egg shell and internal miracidium membrane of Schistosoma japonicum adults, and uses genetic engineering technology to amplify the light chain, The heavy chain variable region gene, and then the heavy chain variable region gene V H wIgG 1 C H1 , light chain variable region gene V L wIgG 1 C L Fd and full-length L were amplified by PCR, and then Fd and full-length L were amplified by overlapping extension PCR to generate a chimeric Fab gene fragment, and the chimeric Fab gene was connected to the vector pComb3XSS to construct a prokaryotic expression vecto...

AI Technical Summary

Problems solved by technology

[0004] At present, the treatment of schistosomiasis is mainly based on praziquantel chemotherapy. Although the treatment of praziquantel can obtain reliable curative effect, the death of acute schistosomiasis infection occurs from time to time. There is no effective treatment method for chronic schistosomiasis liver fibrosis, which can cause significant Portal hypertension, resulting in upper gastrointestinal bleeding, splenomegaly, and ascites, is the main cause of death and loss of labor in patients

In addition, the long-term use of a single chemotherapy drug may cause drug resistance, and long-term use may also induce drug-resistant strains.

Therefore, there are still many problems in the treatment of schistosomiasis, which directly affect the investment environment of the country and the construction of a harmonious society and a well-off society

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