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

Alarcon, Marcelo; FUENTES, EDUARDO; Maldonado, Ximena; Mardones, Claudia; Palomo, Ivan

Abstract

© 2019In 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. • 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).

Más información

Título según SCOPUS: Methodology of generation and purification of anti-beta 2 glycoprotein I antibodies
Título de la Revista: METHODSX
Volumen: 6
Editorial: Elsevier
Fecha de publicación: 2019
Página de inicio: 986
Página final: 992
Idioma: English
DOI:

10.1016/J.MEX.2019.04.023

Notas: WOS-ESCI, SCOPUS