Specificity of recombinant antibodies generated from multiple sclerosis cerebrospinal fluid probed with a random peptide library.
We generated recombinant antibodies (rAbs) from over-represented IgG sequences expressed by single plasma cells from multiple sclerosis (MS) cerebrospinal fluid (CSF). Panning of a phage-displayed random peptide library with the rAbs revealed several specific peptide sequences. Inhibition assays confirmed specific binding of the peptides to the antigen-binding site of the antibody. The native IgG of MS CSF from which the recombinant antibody was cloned also recognized these peptides. Our data demonstrate that MS rAb reflects the specificity of IgG in the CSF. Thus, the epitopes/mimotopes identified by MS rAb may provide clues to disease-relevant antigens.
Yu
Xiaoli
X
Department of Neurology, University of Colorado Health Sciences Center, 4200 E. 9th Avenue, Mail Stop B182, Denver, Colorado 80262, USA. xiaoli.yu@uchsc.edu
Gilden
Donald H
DH
Ritchie
Alanna M
AM
Burgoon
Mark P
MP
Keays
Kathryne M
KM
Owens
Gregory P
GP
D000328:Adult; D000906:Antibodies / Q000134:cerebrospinal fluid*; D000918:Antibody Specificity; D015153:Blotting, Western / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008875:Middle Aged; D009103:Multiple Sclerosis / Q000097:blood / Q000134:cerebrospinal fluid* / Q000276:immunology; D019151:Peptide Library; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
D000906:Antibodies; D000939:Epitopes; D019151:Peptide Library; D012333:RNA, Messenger; D011994:Recombinant Proteins