An ultrasensitive method for the quantitation of active and inactive GLP-1 in human plasma via immunoaffinity
Derek L Chappell1, Anita Yh Lee, Jose Castro-Perez
1Clinical Development Laboratory, Merck Research Laboratories, Rahway, NJ, USA.
Background:
Measuring endogenous levels of incretin hormones, like GLP-1, is critical in the development of antidiabetic compounds. However, the assays used to measure these molecules often have analytical issues.
Results:
We have developed an ultrasensitive, highly-selective immunoaffinity LC-MS/MS (IA LC-MS/MS) assay capable of quantitating endogenous levels of active (7-36 amide) and inactive (9-36 amide) GLP-1 in human plasma. We performed fit-for-purpose validation of the assay by assessing the following assay performance characteristics: inter-assay precision, sensitivity, spike recovery, dilution linearity, absolute recovery, matrix effect, immunoprecipitation efficiency, and food effect.
Conclusion:
We have developed a robust analytical method for the quantitation of endogenous active and inactive GLP-1 in human plasma. In addition, we employed this method to measure the typical changes in GLP-1 levels after food intake. The sensitivity of this assay is better than another LC-MS/MS GLP-1 assay previously reported and many commercially available immunoassays. This important analytical tool could be used to qualify and/or harmonize the different immunoassays used for the quantitation of GLP-1.
