Spot Synthesis: An Optimized Microarray to Detect IgE Epitopes.
Peptide microarrays have become increasingly more affordable in recent years with the SPOT technique being one of the most frequently used methods for synthesis and screening of peptides in arrays. Here, a protocol is presented for the identification of the amino acid sites involved in the conversion of human IgG to IgE response during the passive administration of therapeutic, anti-snake venom sera. Similarly, the minimal region of both the IgG and IgE binding epitopes, important for its interaction with ligand, were identified. As the ratio of concentrations for IgG to IgE in human serum is 1:10,000, also presented is a reproductive protocol of chemiluminescence-scanning for the detection of both immunoglobulins.
FIOCRUZ, Center of Technological Development in Health (CDTS) / National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN), FIOCRUZ, Rio de Janeiro, Brazil. dsimone@cdts.fiocruz.br.
Salvatore G
SG
De-Simone
Department of Cellular andMolecularBiology, Federal FluminenseUniversity,Biology Institute, Niterói, Rio de Janeiro, Brazil.
Paloma
P
Napoleão-Pêgo
IOCRUZ, Center of Technological Development in Health (CDTS) / National Institute of Science and Technology for Innovation on Neglected Diseases (INCT-IDN) FIOCRUZ, Rio de Janeiro, Brazil.
Thatiane S
TS
De-Simone
D000107:Acetylation; D000818:Animals; D002482:Cellulose / Q000737:chemistry; D060326:Chemistry Techniques, Synthetic; D018604:Epitope Mapping / Q000379:methods*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D008567:Membranes, Artificial; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D040081:Protein Array Analysis / Q000379:methods*
D007074:Immunoglobulin G; D008567:Membranes, Artificial; D010455:Peptides; D007073:Immunoglobulin E; D002482:Cellulose
Allergy; Epitope structure; Hypersensitivity; IgE detection; Microarray; Protein–peptide interactions; Spot-synthesis