Development of a mix-and-read assay for human asprosin using antibody–oligonucleotide probes and thermofluorimetric analysis

Abstract

Adipose tissue, or fat tissue, can now be classified as an endocrine organ as it responds to stimuli by secreting a range of hormones, termed adipokines, which regulate the functions of various other tissues and organs. Because novel adipokines continue to be discovered and characterized by researchers, there is an enduring need for the development of new analytical assays that target these hormones. Discovered recently, asprosin is an adipokine hormone secreted by white adipose tissue (WAT) during fasting which has been implicated for its important effects on the liver, skeletal muscle, hypothalamus, pancreas, and possibly other tissues. While standard immunoassays have been developed, the continued surge in research on asprosin's function would greatly benefit from an assay with homogeneous, mix-and-read workflow, and the nanomolar clinical range makes this goal more feasible. In this work, we developed such an assay for asprosin using our thermofluorimetric analysis (TFA) methods with antibody–oligonucleotide conjugate probes. The assay, achievable in less than one hour, was successfully validated by quantifying native levels of asprosin in human serum collected from fasting, nonfasting, type II diabetic, and obese donors.