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Novel and sensitive noncompetitive enzyme immunoassay for peptides



Novel and sensitive noncompetitive enzyme immunoassay for peptides



Biochemical & Biophysical Research Communications 160(1): 40-45



A novel and sensitive noncompetitive enzyme immunoassay for peptides is described (Fig. 1). Peptides were biotinylated using sulfosuccinimidyl-6-(biotinamido)hexanoate and were trapped onto anti-peptide IgG-coated polystyrene balls. After washing the polystyrene balls to eliminate other biotinylated substances, the biotinylated peptides were eluted with HCl and were reacted with anti-peptide Fab'-peroxidase conjugate. The complex formed was trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of angiotensin I as a model peptide was 13 fg (10 amol)/tube and 0.8 ng/l of plasma, which was 80 to 480-fold lower than those previously reported by competitive radioimmunoassay and competitive enzyme immunoassay. And other peptides could also be measured more sensitively by the present noncompetitive enzyme immunoassay method than by competitive immunoassays.

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Accession: 007602581

Download citation: RISBibTeXText

PMID: 2523704

DOI: 10.1016/0006-291x(89)91617-3


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