Expression And Identification Of β₂-GPI In Bac-to-Bac Baculovirus Expression System

The antiphospholipidsyndrome(APS) is a multi-system autoimmune diseasecharacterized by artery/venous thrombosis、 thrombocytopenia and recurrentpregnancy loss. Antiphospholipid antibodies (APA) positive is regarded as thelaboratory diagnosis evidence of APS. β2-GPⅠis a plasma glycoprotein, as theprimary target antigens against antiphospholipid antibodies, played an important rolein the pathogenesis of APS combined with the corresponding antibodies-anti-β2-GPⅠantibodies,.In this study, the insect-baculovirus expression system was used to expressβ2-GPⅠto gain the recombinant virus DNA. After PCR identification, therecombinant virus DNA was transfected into SF9cells to obtain p1venom byliposome method. PCR was performed being virus DNA extracted from infected SF9cell as template. P2, P3venom were collected from cell culture fluid when fluorescentcells increased to a certain degree. Finally, protein expression was performed in SF9cells cultured in serum-free medium by using P3venom. SDS-PAGE preliminaryproved thatβ2-GPⅠ gene was expressed in SF9insect cells.This study lay a foundation for the study ofβ2-GPⅠ’s biological characteristics,physiological functions, especially the function in the process of APS thrombosis.