Production and characterisation of a recombinant scFv reactive with human gastrointestinal carcinomas.

摘要: SC142-reactive antigen are highly glycosylated glycoproteins expressed on tissues of gastric and colon cancers but not normal tissues. Murine SC142 antibody specific for the has been produced by immunisation with SNU16 stomach cancer cells. However, several potential problems such as high immunogenicity poor tumour penetration owing to their large size. To improve in vivo, recombinant single-chain fragments have using original hybridoma cells a source variable heavy- light-chain-encoding genes. The use polymerase chain reaction, expression cloning technology gene systems E. coli led production fragments, which was similar activity parent confirmed immunohistochemistry. Analysis DNA sequencing, SDS–PAGE Western blotting demonstrated integrity fragments. Competitive ELISA showed that originated from antibody. BIAcore biosensor binding experiments had an ideal dissociation rate constant imaging reagent. These results illustrate application these novel products immunodiagnostic further immunotherapeutic reagent. British Journal Cancer (2002) 87, 405–413. doi:10.1038/sj.bjc.6600365 www.bjcancer.com © 2002 Research UK