Anti-schistosomiasis monoclonal antibody NP11-4 single-chain antibody, preparation and use thereof

Abstract

The invention discloses a single-chain antibody of an anti-schistosome monoclonal NP11-4 as well as a preparation method and application thereof, and relates to the field of gene engineering and the field of therapeutic drugs against schistosomiasis. The invention is to use a monoclonal antibody NP11-4 located at adult membrane, cercaria memberane, schistosomulum membrane, egg shell and miracidium in egg of Schistosoma japonicum, extract total RNA of hybridoma cells through a gene engineering technology, adopt RT-PCR for proliferation of genes VH and VL, and use overlap-extension PCR to connect the genes VH and VL into a single-chain antibody gene in the form of VH<-linker-VL, which is subsequently connected with a pBAD / gIIIA carrier and cloned into Top10F'of Escherichia Coli to induce a soluble expression of the target gene. The zone from amino acid No.1 to No. 122 of the expressed single-chain antibody of the anti-schistosome monoclonal NP11-4 is a heavy chain gene repertoire of the single-chain antibody (VH), and the zone from the amino acid No.141 to No.254 is a light chain gene repertoire (VL) of the single-chain antibody, wherein the linker between the heavy and light chain gene repertoires consists of 18 amino acids including repeated glycines and serines, and the single-chain antibody has 254 amino acids in full length.

Description

technical field [0001] The invention relates to an anti-schistosomiasis monoclonal antibody NP11-4 single-chain antibody and its preparation method and application, belonging to the genetic engineering technology for preparing the anti-schistosomiasis single-chain antibody. The single-chain antibody 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. The light chain and heavy chain can be amplified by genetic engineering technology. Variable region gene, with V H -linker-V L The form is connected with the prokaryotic expression vector to prepare single-chain antibody, which is used for the treatment of schistosomiasis or the preparation of anti-schistosomiasis immunotoxin. The invention relates to the fields of genetic engineering technology and medicine for treating schistosomiasis. Background technique [0002] Schistosomiasis ...

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, death from acute schistosomiasis infection occurs from time to time. There is no effective treatment method for chronic schistosomiasis liver fibrosis, which can cause significant damage. 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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