Expression, production and renaturation of a functional single-chain variable antibody fragment (scFv) against human intercellular adhesion molecule-1 (ICAM-1)

  • Hong Sun
  • Yu Ding
  • Songling Yang
  • Zuoqun Yin
  • Yuhuan Yue

Abstract

The single-chain variable antibody fragment (scFv) against human intercellular adhesion molecule-1 (ICAM-1) was expressed at a high level in Escherichia coli as inclusion bodies. We attempted to refold the scFv by ion-exchange chromatography (IEC), dialysis and dilution. The results show that the column chromatography refolding by Q Sepharose high performance (Q HP) had remarkable advantages over the conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield was higher by this method, which is about 60 mg/l. The purity of the final product was greater than 90%, as shown by denaturing gel electrophoresis. Enzyme-linked immunosorbent assay (ELISA), cell culture and animal experiments were used to assess the immunologic properties and biologic activities of the renatured scFv.

Keywords: Intercellular adhesion molecule-1, single-chain variable antibody fragment, expression, purification, renaturation, biological activity.

African Journal of Biotechnology, Vol 13(14), 1588-1596

Author Biographies

Hong Sun
Department of Biological Pharmacy, Heilongjiang Vocational College of Biology Science and Technology, Harbin 150025, PR, China
Yu Ding
Laboratory Animal Center, Jilin University, Changchun, 130062, PR China
Songling Yang
Department of Biological Pharmacy, Heilongjiang Vocational College of Biology Science and Technology, Harbin 150025, PR, China
Zuoqun Yin
Department of Biological Pharmacy, Heilongjiang Vocational College of Biology Science and Technology, Harbin 150025, PR, China
Yuhuan Yue
Institute of Military Veterinary, Academy of Military Medical Sciences of the Chinese People's Liberation Army, Changchum, 130062, PR China
Published
2015-06-25
Section
Articles

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eISSN: 1684-5315