Rapid and efficient identification of epitopes/mimotopes from random peptide libraries.
Phage-displayed random peptide libraries are important tools in identifying novel epitopes/mimotopes that may lead to the determination of antigen specificity. In this approach, high-affinity phage peptides are enriched by affinity selection (panning) on a monoclonal antibody. To facilitate identification of all potential phage peptides specific for recombinant monoclonal antibodies (rAbs) previously generated from clonally expanded plasma cells from the cerebrospinal fluid of patients with multiple sclerosis (MS), we developed a high-throughput method to determine phage specificity. In contrast to the 8-9 days needed in the standard large-scale method of amplifying phage clones for ELISA, the high-throughput method takes only 1 day. ELISA using phage clones amplified directly in 96-well plates avoids large-scale phage purification and enables rapid identification of specific epitopes/mimotopes. This technique will expedite identification of MS-specific peptides that can be used to discover the corresponding protein antigens.
Yu
Xiaoli
X
Department of Neurology, University of Colorado Health Sciences Center, 4200 East 9th Avenue, Mail Stop B182, Denver, CO 80262, USA.
Owens
Gregory P
GP
Gilden
Donald H
DH
D000595:Amino Acid Sequence; D017104:Bacteriophage M13; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D009103:Multiple Sclerosis / Q000134:cerebrospinal fluid / Q000276:immunology; D019151:Peptide Library; D011994:Recombinant Proteins / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D000939:Epitopes; D019151:Peptide Library; D011994:Recombinant Proteins; D004247:DNA