Highly specific novel method for isolation and purification of lupus anti-DNA antibody via oligo-(dT) magnetic beads.
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.
Pavlovic
Mirjana
M
Department of Biological Sciences, Charles E. Schmidt College of Science, Florida Atlantic University, Boca Raton, Florida 33451, USA. pmirjana@aol.com
Chen
Ran
R
Kats
Anna M
AM
Cavallo
Michelle F
MF
Saccocio
Seth
S
Keating
Patricia
P
Hartmann
James X
JX
D000974:Antibodies, Antinuclear / Q000302:isolation & purification*; D000915:Antibody Affinity; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008180:Lupus Erythematosus, Systemic / Q000175:diagnosis*; D009838:Oligodeoxyribonucleotides; D012680:Sensitivity and Specificity
D000974:Antibodies, Antinuclear; D009838:Oligodeoxyribonucleotides; C027903:oligo (dT)