Low-Cost Peptide Microarrays for Mapping Continuous Antibody Epitopes.
With the increasing need for understanding antibody specificity in antibody and vaccine research, pepscan assays provide a rapid method for mapping and profiling antibody responses to continuous epitopes. We have developed a relatively low-cost method to generate peptide microarray slides for studying antibody binding. Using a setup of an IntavisAG MultiPep RS peptide synthesizer, a Digilab MicroGrid II 600 microarray printer robot, and an InnoScan 1100 AL scanner, the method allows the interrogation of up to 1536 overlapping, alanine-scanning, and mutant peptides derived from the target antigens. Each peptide is tagged with a polyethylene glycol aminooxy terminus to improve peptide solubility, orientation, and conjugation efficiency to the slide surface.
DNA Array Core, Department of Immunology and Microbial Science, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA, USA.
Ryan
R
McBride
DNA Array Core, Department of Immunology and Microbial Science, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA, USA.
Steven R
SR
Head
Protein and Nucleic Acid CoreFacility, Department ofImmunology and Microbial Science,, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA, USA.
Phillip
P
Ordoukhanian
DNA Array Core, Department of Immunology and Microbial Science, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA, USA. mlaw@scripps.edu.
Mansun
M
Law
D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D002482:Cellulose / Q000737:chemistry; D018604:Epitope Mapping / Q000191:economics*; D005449:Fluorenes / Q000737:chemistry; D006801:Humans; D055767:Immobilized Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011327:Printing; D040081:Protein Array Analysis / Q000191:economics*
C496789:9-fluorenylmethoxycarbonyl; D000906:Antibodies; D005449:Fluorenes; D055767:Immobilized Proteins; D008567:Membranes, Artificial; D010455:Peptides; D002482:Cellulose; D000409:Alanine
Aminooxy-tagged peptide; Antibody epitope; Continuous epitope; Epitope map ping; Linear epitope; Pepscan; Peptide array; Peptide microarray