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 Citations (Scopus)

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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Single-Chain Antibodies
Escherichia coli
Epitopes
Affinity chromatography
Monoclonal antibodies
Antigens
Assays
Fusion reactions
Proteins
Glutathione Transferase
Affinity Chromatography
Histidine
Thrombin
Sepharose
Glutathione
Digestion
Western Blotting
Monoclonal Antibodies
Neoplasms

ASJC Scopus subject areas

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

Cite this

Expression of recombinant Lym-I single-chain Fv in Escherichia coli. / Song, Kyung Bin; Won, Misun; Meares, Claude F.

In: Biotechnology and Applied Biochemistry, Vol. 28, No. 2, 1998, p. 163-167.

Research output: Contribution to journalArticle

Song, KB, Won, M & Meares, CF 1998, 'Expression of recombinant Lym-I single-chain Fv in Escherichia coli', Biotechnology and Applied Biochemistry, vol. 28, no. 2, pp. 163-167.
Song, Kyung Bin ; Won, Misun ; Meares, Claude F. / Expression of recombinant Lym-I single-chain Fv in Escherichia coli. In: Biotechnology and Applied Biochemistry. 1998 ; Vol. 28, No. 2. pp. 163-167.
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