Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system

M. Igarashi, E. Matsuura, Y. Igarashi, H. Nagae, Y. Matsuura, K. Ichikawa, T. Yasuda, D. R. Voelker, T. Koike

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A full-length cDNA coding a human β2-glycoprotein I (β2-GPI) was introduced into the baculovirus genome to construct a recombinant baculovirus. Spodoptera frugiperda (Sf9) cells were infected with the recombinant baculovirus. A protein (mol. wt 43000) reactive with anti-β2-GPI antisera was produced in the insect cells and secreted into the culture medium. The recombinant β2-GPI was purified from the culture supernatant by sequential cardiolipin (CL)-affinity column chromatography and gel filtration. The N-terminal amino acid sequence of the protein was identical to that of the native β2-GPI purified from human sera, and a putative signal peptide was cleaved from the secreted form of the recombinant protein. The purified recombinant protein had a cofactor activity which enhances CL binding of anticardiolipin antibodies (aCL) in systemic lupus erythematosus (SLE) patients, as well as the native β2-GPI. Thus, the β2-GPI expressed in insect cells is an immunologically active cofactor.

Original languageEnglish
Pages (from-to)19-25
Number of pages7
JournalClinical and Experimental Immunology
Volume93
Issue number1
Publication statusPublished - Jan 1 1993
Externally publishedYes

Keywords

  • anticardiolipin antibodies
  • baculovirus expression
  • glycosylation
  • recombinant DNA

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Igarashi, M., Matsuura, E., Igarashi, Y., Nagae, H., Matsuura, Y., Ichikawa, K., Yasuda, T., Voelker, D. R., & Koike, T. (1993). Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system. Clinical and Experimental Immunology, 93(1), 19-25.