Expression of recombinant Lym-I single-chain Fv in Escherichia coli

Kyung Bin Song, Misun Won, Claude F. Meares

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Lym-I single-chain Fv (sFv) can be used for targeted radiodiagnosis and therapy of B-lymphocytic malignancies. Lym-I sFv was constructed and expressed as a glutathione S-transferase fusion protein, using a (G4S)3 linker connecting the C-terminus of the V(H) domain and the N-terminus of the V(L) domain of Lym-I. Six histidine residues and an E Tag epitope were introduced at the C-terminus of the sFv. Lym-I sFv was purified with glutathione-Sepharose 4B affinity chromatography followed by digestion with thrombin. Lym-I sFv of 28 kDa was confirmed by Western blotting with anti-(E Tag) monoclonal antibody. An antigen binding assay of Lym-I and a CD study indicated that it is functionally active.

Original languageEnglish (US)
Pages (from-to)163-167
Number of pages5
JournalBiotechnology and Applied Biochemistry
Volume28
Issue number2
StatePublished - 1998

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology

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