A sensitive, robust high-throughput electrochemiluminescence assay for rat insulin.
In diabetes research, mouse and rat models are used for in vivo experiments, and quantification of insulin in serum samples under different pathophysiological conditions and after treatment with compounds is essential. There are few commercial radioimmunoassay and enzyme-linked immunosorbent assay (ELISA) assay kits to determine the rat/mouse plasma levels of insulin. However, reliability in insulin measurements using the available biological assays is a great concern. The authors report a robust, extremely sensitive electrochemiluminescence (ECL) insulin assay using the Origen technology platform. The assay performance, as judged by the Z' value of 0.82+/-0.03 and the signal-to-background (S/B) ratio of 133, suggests that this is a robust and reliable assay. The intra-assay and interassay variation is less than 5%. The dynamic range of detection for insulin is 5 pg to 5 ng in the ECL assays. Recovery of insulin was about 100% when different volumes of serum were spiked with exogenous insulin. These results suggest that the ECL insulin assay is an extremely sensitive, robust, nonradioactive homogeneous assay and can be used successfully to determine the insulin levels in rodent serum samples.
Golla
Rajasree
R
Drug Discovery, Pharmaceutical Research Institute, Bristol-Myers Squibb, Princeton, NJ 08543-5400, USA.
Seethala
Ramakrishna
R
D000818:Animals; D000911:Antibodies, Monoclonal; D002138:Calibration; D004563:Electrochemistry / Q000379:methods*; D007328:Insulin / Q000097:blood* / Q000276:immunology; D008163:Luminescent Measurements; D051379:Mice; D051381:Rats; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D007328:Insulin