A Biotin-avidin-based Enzyme Immunoassay for Beta H-endorphin

Overview

Journal Pharm Res

Specialties Pharmacology
Pharmacy

Date 1988 Apr 1

PMID 2977649

Citations 2

Authors

G Hochhaus

W Sadee

Affiliations

Soon will be listed here.

Abstract

An avidin-biotin enzyme-linked immunosorbent assay (ELISA) is described for beta h-endorphin (beta h-EP). Microtiter plates coated with commercially available antibodies were used together with beta h-EP tracer derivatives that were biotinylated in positions 24, 28, and 29 via a C6 spacer arm. Nonspecific binding of biotinylated derivatives to the microtiter plates was blocked with a mixture of 1% casein and 10% ethanolamine in 0.1 M NaHCO3. A sequential saturation procedure using a high-affinity antiserum in combination with an avidin-alkaline phosphatase complex matched the sensitivity of reported radioimmunoassays (RIAs), with a detection limit of 0.5 fmol/assay. The intra- and interassay coefficients of variation were 5 and 12%, respectively. Results obtained by ELISA and RIA showed good correlations (r = 0.95). The beta-EP concentration in extracted rat plasma after high-performance liquid chromatographic (HPLC) fractionation was determined by this method to be 1600 fmol/mol.

Citing Articles

A fast universal immobilization of immunoglobulin G at 4 °C for the development of array-based immunoassays.

Guo S, Chen P, Chen M, Cheng Y, Lin J, Lee H PLoS One. 2012; 7(12):e51370.

PMID: 23236488 PMC: 3517563. DOI: 10.1371/journal.pone.0051370.

[Biocytin13]dynorphin A 1-13 amide: a potential probe for the kappa-opioid receptor.

Hochhaus G, Patthy A, Schwietert R, Santi D, Sadee W Pharm Res. 1988; 5(12):790-4.

PMID: 2907785 DOI: 10.1023/a:1015944903360.

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