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

M. Igarashi, Eiji 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 - 1993
Externally publishedYes

Fingerprint

beta 2-Glycoprotein I
Baculoviridae
Insects
Glycoproteins
Cardiolipins
Recombinant Proteins
Sf9 Cells
Spodoptera
Anticardiolipin Antibodies
Protein Sorting Signals
Affinity Chromatography
Systemic Lupus Erythematosus
Gel Chromatography
Culture Media
Immune Sera
Amino Acid Sequence
Proteins
Complementary DNA
Genome
Serum

Keywords

  • anticardiolipin antibodies
  • baculovirus expression
  • glycosylation
  • recombinant DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system. / Igarashi, M.; Matsuura, Eiji; Igarashi, Y.; Nagae, H.; Matsuura, Y.; Ichikawa, K.; Yasuda, T.; Voelker, D. R.; Koike, T.

In: Clinical and Experimental Immunology, Vol. 93, No. 1, 1993, p. 19-25.

Research output: Contribution to journalArticle

Igarashi, M, Matsuura, E, Igarashi, Y, Nagae, H, Matsuura, Y, Ichikawa, K, Yasuda, T, Voelker, DR & Koike, T 1993, 'Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system', Clinical and Experimental Immunology, vol. 93, no. 1, pp. 19-25.
Igarashi, M. ; Matsuura, Eiji ; Igarashi, Y. ; Nagae, H. ; Matsuura, Y. ; Ichikawa, K. ; Yasuda, T. ; Voelker, D. R. ; Koike, T. / Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system. In: Clinical and Experimental Immunology. 1993 ; Vol. 93, No. 1. pp. 19-25.
@article{c6be2758000049e481a493bb9932f39e,
title = "Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system",
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.",
keywords = "anticardiolipin antibodies, baculovirus expression, glycosylation, recombinant DNA",
author = "M. Igarashi and Eiji Matsuura and Y. Igarashi and H. Nagae and Y. Matsuura and K. Ichikawa and T. Yasuda and Voelker, {D. R.} and T. Koike",
year = "1993",
language = "English",
volume = "93",
pages = "19--25",
journal = "Clinical and Experimental Immunology",
issn = "0009-9104",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Expression of anticardiolipin cofactor, human β2-glycoprotein I, by a recombinant baculovirus/insect cell system

AU - Igarashi, M.

AU - Matsuura, Eiji

AU - Igarashi, Y.

AU - Nagae, H.

AU - Matsuura, Y.

AU - Ichikawa, K.

AU - Yasuda, T.

AU - Voelker, D. R.

AU - Koike, T.

PY - 1993

Y1 - 1993

N2 - 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.

AB - 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.

KW - anticardiolipin antibodies

KW - baculovirus expression

KW - glycosylation

KW - recombinant DNA

UR - http://www.scopus.com/inward/record.url?scp=0027172725&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027172725&partnerID=8YFLogxK

M3 - Article

C2 - 8324900

AN - SCOPUS:0027172725

VL - 93

SP - 19

EP - 25

JO - Clinical and Experimental Immunology

JF - Clinical and Experimental Immunology

SN - 0009-9104

IS - 1

ER -