Monoclonal antiidiotypic antibody NP30 single-specificity double-chain antibody of Japanese blood fluke, preparation method and uses thereof

A technology of idiotype antibody and double-chain antibody, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve problems affecting the construction of a harmonious society and a well-off society, losing labor force, and affecting the national investment environment, etc.

Inactive Publication Date: 2009-02-18
NANJING MEDICAL UNIV
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Problems solved by technology

[0003] 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, long-term application of praziquantel may 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

Method used

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  • Monoclonal antiidiotypic antibody NP30 single-specificity double-chain antibody of Japanese blood fluke, preparation method and uses thereof
  • Monoclonal antiidiotypic antibody NP30 single-specificity double-chain antibody of Japanese blood fluke, preparation method and uses thereof
  • Monoclonal antiidiotypic antibody NP30 single-specificity double-chain antibody of Japanese blood fluke, preparation method and uses thereof

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Embodiment Construction

[0067] Detailed description of the embodiment of the present invention:

[0068] Expression of monoclonal anti-idiotypic antibody NP30 monospecific diabody against Schistosoma japonicum

[0069] Application of overlap PCR method to amplify diabody gene V H -linker-V L(Diabody, referred to as D), the gene and the pBAD / gIIIA vector were digested with SacI and EcoRI, and connected to construct the recombinant plasmid pBAD / gIIIA-D. The recombinant plasmid was transformed into E. coli TOP10F' to express NP30 monospecific diabody. SDS-PAGE and Westemblot identification results show that the protein ratio exists in two forms of inclusion body and soluble.

[0070] Materials and Methods

[0071] 1. Strain and plasmid: E.coli TOP10F', preserved in this laboratory. pMD18-T-V H , pMD18-T-V L Plasmids, in which the NP30 variable region light chain (VL) and heavy chain (VH) genes are respectively cloned, and the pBAD / gIII plasmid is kept in this laboratory.

[0072] 2. Tool enzymes...

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Abstract

The present invention relates to a schistosoma japonicum monoclonal anti-idiotypic antibody NP30 monospecific diabody and a preparation method and applications thereof., which belong to the field of genetic engineering technology and immunotherapy drug preparation. The preparation method utilizes the genetic engineering technology to prepare the schistosoma japonicum monoclonal anti-idiotypic antibody NP30 monospecific diabody and adopts overlap PCR to amplify diabody gene VH-linker-VL (diabody, D for short), the length of linker chooses five amino acid residues, the sequence is GGGGS, and two scFv molecules are forced to mutually form VH-VL pairing, and bound together by a non-covalent bond to form a dipolymer, which is called diabody. The diabody can be used to treat acute schistosome infection and immunoprotection against schistosomiasis.

Description

technical field [0001] The present invention relates to a Schistosoma japonicum monoclonal anti-idiotypic antibody NP30 monospecific diabody and its preparation method and application. , using the overlap PCR method to amplify the diabody gene V H -linker-V L (Diabody, referred to as D), the length of the linker is 5 amino acid residues, and the sequence is GGGGS, forcing two scFv molecules to form a V H and V L Paired, non-covalently bonded dimers are known as diabodies. The antibody can be used for the treatment of acute schistosomiasis infection and immune protection of schistosomiasis. Background technique [0002] The schistosomes that harm the human body mainly include Schistosoma japonicum, Schistosoma haematobium and Schistosoma mansoni, which are mainly distributed in Asia (east and south), Africa and South America, and the number of patients is about 200 million. The epidemic in my country is Japan Schistosomiasis is distributed in the Yangtze River Basin and 1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N15/13C12N15/70G01N33/577A61K39/395A61P33/12
CPCY02A50/30
Inventor 朱进朱晓娟许静唐奇冯振卿管晓虹
Owner NANJING MEDICAL UNIV
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