Kouts S, Bunn C L, Steinkasserer A, Krilis S
Dept. Immunology, Allergy and Infectious Disease, St. George Hospital, University of N.S.W., Australia.
FEBS Lett. 1993 Jul 12;326(1-3):105-8. doi: 10.1016/0014-5793(93)81771-q.
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.
为了能够合成β2-糖蛋白I突变体，我们建立了一个稳定的中国仓鼠卵巢细胞系，该细胞系每天可表达高达2.9微克/10⁶个细胞的人β2-糖蛋白I。在改良的抗心磷脂ELISA中，重组β2-糖蛋白I的辅因子活性与纯化的天然蛋白相同。自身免疫型抗心磷脂抗体以剂量依赖的方式需要重组β2-糖蛋白I来结合心磷脂，而感染型抗体的结合则受到抑制。在无血清培养基中纯化的重组β2-糖蛋白I以两种寡糖形式存在，去糖基化后其表观分子量与去糖基化的天然蛋白相同。
