Methodology of generation and purification of anti-beta 2 glycoprotein I antibodies

摘要

class="kwd-title">Method name: Generation and purification of Anti-Beta 2 Glycoprotein I antibodies class="kwd-title">Keywords: Beta 2 glycoprotein I, Antibodies, Purification, Antiphospholipid syndrome class="head no_bottom_margin" id="abs0010title">AbstractIn this Method Article we are showing the methodology for generation and purification of Anti-Beta 2 Glycoprotein I (β2GPI) antibodies. First β2GPI was purified from human plasma, and recognized by Western Blot and anti-β2GPI antibodies of serum from patients with antiphospholipid syndrome (APS). The C57BL/6 mice were immunized intraperitonealy with 150 μg of protein in adjuvant (β2GPI or bovine serum albumin) on days 1, 8 and 14. Then the anti-β2GPI antibodies were purified by affinity chromatography (Affi-Gel protein A sepharose) and affinity column using human β2GPI coupled to CNBr-activated Sepharose 4B. Titles of anti-β2GPI antibodies were determined by ELISA assays. class="first-line-outdent" id="lis0005">

• We purified β2GPI with great efficacy and that is recognized antigenically by serum from patients with SAP or an anti-β2gpi antibody.

• We found that our purified antibody had 13 fold increased activity in ELISA test compared with the control and in Western Blot recognized with β2GPI (reference and purified).