Enzyme-linked immunosorbent assay for an octapeptide based on a genetically engineered fusion protein.
Traditional chemical means of preparing enzyme-ligand conjugates for use in enzyme-linked immunosorbent assays (ELISAs) lead to the production of multisubstituted enzyme-ligand conjugates with a high degree of variability in the site of ligand attachment. A genetically engineered fusion protein was prepared in order to investigate the feasibility of controlled production of conjugates for use in ELISAs. Specifically, a synthetic octapeptide was fused with bacterial alkaline phosphatase. The resulting enzyme-peptide conjugate is monosubstituted (one peptide per subunit), has a single site of attachment, and results in assays with good response characteristics. The use of such fusion proteins, which combine small analyte peptides with enzyme labels, can lead to a new approach to improved assays for numerous biomolecules, including peptide pharmaceuticals, neurotransmitters, hormones, cell surface antigens, etc.
Department of Chemistry, University of Kentucky, Lexington 40506-0055.
A
A
Witkowski
S
S
Daunert
M S
MS
Kindy
L G
LG
Bachas
D000469:Alkaline Phosphatase / Q000737:chemistry; D000595:Amino Acid Sequence; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology; D005818:Genetic Engineering; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis* / Q000138:chemical synthesis; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase