Highly Specific Novel Method for Isolation and Purification of Lupus Anti-DNA Antibody via Oligo-(dT) Magnetic Beads

Authors: PAVLOVIC, MIRJANA; CHEN, RAN; KATS, ANNA M.; CAVALLO, MICHELLE F.; SACCOCIO, SETH; KEATING, PATRICIA; HARTMANN, JAMES X.

Source: Annals of the New York Academy of Sciences, Volume 1108, Number 1, June 2007 , pp. 203-217(15)

Publisher: Wiley-Blackwell

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Abstract:

A novel method for isolation and purification of anti-ssDNA antibodies from human sera is developed. The process involves: antibody purification based on their affinity for single-stranded sequence of thymidines and removal of remaining components via protein G coated magnetic beads, with high affinity for only IgG subclass. The high degree of purity and molecular weights of healthy versus lupus anti-ssDNA antibodies were confirmed by SDS-PAGE and silver staining. Western blot confirmed IgG isotype. This novel technique allows for diagnostic purposes, structural and functional analysis of anti-DNA antibodies, and studies of their role in autoimmune diseases.

Keywords: systemic lupus erythematosus (SLE); anti-ssDNA antibody purification; magnetic beads; affinity chromatography; SDS-PAGE; silver staining; recovery rate; antibody hydrolytic activity

Document Type: Research article

DOI: http://dx.doi.org/10.1196/annals.1422.023

Affiliations: 1: Department of Biological Sciences, Charles E. Schmidt College of Science, Florida Atlantic University, Boca Raton, Florida, USA

Publication date: 2007-06-01