Pavlovic Mirjana, Chen Ran, Kats Anna M, Cavallo Michelle F, Saccocio Seth, Keating Patricia, Hartmann James X
Department of Biological Sciences, Charles E. Schmidt College of Science, Florida Atlantic University, Boca Raton, Florida 33451, USA.
Ann N Y Acad Sci. 2007 Jun;1108:203-17. doi: 10.1196/annals.1422.023.
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.