Identification of measles virus epitopes using an ultra-fast method of panning phage-displayed random peptide libraries.
Phage-displayed random peptide libraries, in which high affinity phage peptides are enriched by repetitive selection (panning) on target antibody, provide a unique tool for identifying antigen specificity. This paper describes a new panning method that enables selection of peptides in 1 day as compared to about 6 days required in traditional panning to identify virus-specific epitopes. The method, termed ultra-fast selection of peptide (UFSP), utilizes phage produced by bacterial infection (phage amplification) directly for subsequent panning. Phage amplified in less than 1h of infection in Escherichia coli are used for binding to target antibody pre-coated in the same wells of an ELISA plate, obviating the need for traditional large-scale amplification and purification. Importantly, phage elution at 37 degrees C was superior to that at room temperature, and phage amplification in a 150-microl volume of E. coli cells was superior to that in 250-microl volume. Application of UFSP to two monoclonal antibodies generated from clonally expanded plasma cells in subacute sclerosing panencephalitis (SSPE) brain identified high-affinity measles virus-specific-peptide epitopes. The UFSP panning methodology will expedite identification of peptides reacting with antibodies generated in other diseases of unknown antigenic specificity such as multiple sclerosis (MS), sarcoidosis and Behcet's disease.
Yu
Xiaoli
X
Department of Neurology, University of Colorado Denver School of Medicine, 4200 East Ninth Avenue, Mail Stop B182, Denver, CO 80262, USA.
Barmina
Olga
O
Burgoon
Mark
M
Gilden
Don
D
D000914:Antibodies, Viral; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000302:isolation & purification*; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology; D008459:Measles virus / Q000737:chemistry*; D019151:Peptide Library
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D019151:Peptide Library