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

D. J. Kim, J. H. Chung, Y. S. Ryu, J. H. Rhim, C. W. Kim, Yousin Suh, H. K. Chung

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

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

Original languageEnglish (US)
Pages (from-to)405-413
Number of pages9
JournalBritish Journal of Cancer
Volume87
Issue number4
DOIs
StatePublished - Aug 12 2002
Externally publishedYes

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Carcinoma
Antibodies
Stomach Neoplasms
Antigens
Neoplasms
Hybridomas
Biosensing Techniques
DNA Sequence Analysis
Colonic Neoplasms
Organism Cloning
Polyacrylamide Gel Electrophoresis
Immunization
Glycoproteins
Western Blotting
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Escherichia coli
Technology
Gene Expression
Light

Keywords

  • Expression cloning
  • SC142 antibody
  • SC142 scFv
  • SC142-reactive antigen

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Kim, D. J., Chung, J. H., Ryu, Y. S., Rhim, J. H., Kim, C. W., Suh, Y., & Chung, H. K. (2002). Production and characterisation of a recombinant scFv reactive with human gastrointestinal carcinomas. British Journal of Cancer, 87(4), 405-413. https://doi.org/10.1038/sj.bjc.6600365

Production and characterisation of a recombinant scFv reactive with human gastrointestinal carcinomas. / Kim, D. J.; Chung, J. H.; Ryu, Y. S.; Rhim, J. H.; Kim, C. W.; Suh, Yousin; Chung, H. K.

In: British Journal of Cancer, Vol. 87, No. 4, 12.08.2002, p. 405-413.

Research output: Contribution to journalArticle

Kim, D. J. ; Chung, J. H. ; Ryu, Y. S. ; Rhim, J. H. ; Kim, C. W. ; Suh, Yousin ; Chung, H. K. / Production and characterisation of a recombinant scFv reactive with human gastrointestinal carcinomas. In: British Journal of Cancer. 2002 ; Vol. 87, No. 4. pp. 405-413.
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