PMID: 25744848Mar 7, 2015Paper

Preparation and application of a polyclonal antibody against the peptides of β₂glycoprotein 1

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Hongxiang XieLongfei Xia

Abstract

To prepare a polyclonal antibody against human and murine β2 glycoprotein 1 (β2GP1) antigen with chemically synthesized β2GP1 peptides, and identify its specificity and pathogenicity. The peptides from the NH₂-terminal 35th-51th amino acids of β₂GP1 were synthesized by standard Fmoc assay, and then used to immunize New Zealand white rabbits after coupling with keyhole limpet hemocyanin (KLH). The polyclonal antibody in the rabbit sera was purified by protein G column. The titer and specificity of the polyclonal antibody were determined by ELISA and Western blot analysis. The total RNA was extracted and the protein lysates were collected from C3H/HeN mouse peritoneal macrophages treated with the above anti-β₂GP1 peptides antibody/β₂GP1 complexes in vitro. And the tissue factor (TF) mRNA and protein expression in the peritoneal macrophages were detected by real-time quantitative PCR and Western blotting. The activation of p38 and NF-κB p65 induced by anti-β₂GP1 peptides antibody/β₂GP1 complexes was determined by Western blotting using phosphor-specific antibodies. Experimental antiphospholipid antibody syndrome (EAPS) mouse model was established in C3H/HeN mice by intraperitoneal injection of anti-β₂GP1 peptides antibody in vivo....Continue Reading

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