Expression of human recombinant beta 2-glycoprotein I with anticardiolipin antibody cofactor activity.
To enable the synthesis of beta 2-glycoprotein I mutants we have established a stable Chinese hamster ovary cell line that expresses human beta 2-glycoprotein I up to 2.9 micrograms/10(6) cells/day. Recombinant beta 2-glycoprotein I is identical to the purified native protein with respect to cofactor activity revealed in a modified anti-cardiolipin ELISA. Autoimmune type anti-cardiolipin antibody requires recombinant beta 2-glycoprotein I in a dose-dependent manner to bind cardiolipin whilst binding of infectious type antibody is inhibited. The purified recombinant beta 2-glycoprotein I in serum free medium exists as two oligosaccharide species which upon deglycosylation have identical apparent molecular weight to the deglycosylated native protein.
Kouts
S
S
Dept. Immunology, Allergy and Infectious Disease, St. George Hospital, University of N.S.W., Australia.
Bunn
C L
CL
Steinkasserer
A
A
Krilis
S
S
D000818:Animals; D017153:Antibodies, Anticardiolipin / Q000302:isolation & purification / Q000378:metabolism*; D001053:Apolipoproteins; D016466:CHO Cells / Q000378:metabolism; D002308:Cardiolipins / Q000276:immunology*; D002845:Chromatography; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D017421:Sequence Analysis; D014162:Transfection; D053482:beta 2-Glycoprotein I
D017153:Antibodies, Anticardiolipin; D001053:Apolipoproteins; D002308:Cardiolipins; D006023:Glycoproteins; D011994:Recombinant Proteins; D053482:beta 2-Glycoprotein I; D004247:DNA