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Biotechnology and Applied Biochemistry (1998) 28, (163–167) (Printed in Great Britain)
Expression of recombinant Lym-1 single-chain Fv in Escherichia coli
Kyung Bin Song*1, Misun Won† and Claude F. Meares‡
*Department of Food Science and Technology, Chungnam National University, Taejon 305-764, Korea, †Cell Cycle and Signal Transduction Research Unit, Korea Research Institute of Bioscience and Biotechnology, Taejon 305-600, Korea, and ‡Department of Chemistry, University of California, Davis, CA 95616, U.S.A.
Abbreviations used: GST, glutathione S-transferase; IPTG, isopropyl b-D-thiogalactoside; mAb, monoclonal antibody; sFv, single-chain Fv. 1 To whom correspondence should be addressed. 
Lym-1 single-chain Fv (sFv) can be used for targeted radiodiagnosis and therapy of B-lymphocytic malignancies. Lym-1 sFv was constructed and expressed as a glutathione S-transferase fusion protein, using a (G4S)3 linker connecting the C-terminus of the VH domain and the N-terminus of the VL domain of Lym-1. Six histidine residues and an E Tag epitope were introduced at the C-terminus of the sFv. Lym-1 sFv was purified with glutathione–Sepharose 4B affinity chromatography followed by digestion with thrombin. Lym-1 sFv of 28 kDa was confirmed by Western blotting with anti-(E Tag) monoclonal antibody. An antigen binding assay of Lym-1 and a CD study indicated that it is functionally active.
Received 15 April 1998/19 May 1998; accepted 22 May 1998
Portland Press Ltd © 1998 |
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