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A study to assess the cross‐reactivity of cellulose membrane‐bound peptides with detection systems: an analysis at the amino acid level
Author(s) -
Mahrenholz Carsten C.,
Tapia Victor,
Stigler Rolf D.,
Volkmer Rudolf
Publication year - 2010
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.1237
Subject(s) - analyte , peptide , chemistry , streptavidin , amino acid , combinatorial chemistry , thiourea , peptide sequence , chromatography , biochemistry , biotin , organic chemistry , gene
The growing demand for binding assays to study protein–protein interaction can be addressed by peptide array‐based methods. The SPOT technique is a widespread peptide‐array technology, which is able to distinguish semi‐quantitatively the binding affinities of peptides to defined protein targets within one array. The quality of an assay system used for probing peptide arrays depends on the well‐balanced combination of screening and read‐out methods. The former address the steady‐state of analyte capture, whereas the latter provide the means to detect captured analyte. In all cases, however, false‐positive results can occur when challenging a peptide array with analyte or detecting captured analyte with label conjugates. Little is known about the cross‐reactivity of peptides with the detection agents. Here, we describe at the amino acid level the potential of (i) 5‐(and 6)‐carboxytetramethylrhodamine (5(6)‐TAMRA), (ii) fluoresceinisothiocyanate in form of the peptide‐bound fluorescein‐substituted thiourea derivative (FITC), and (iii) biotin/streptavidin‐POD to cross‐react with individual amino acids in a peptide sequence. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd.