Potential use of Cypridina luciferin for quantifying alpha 1-acid glycoprotein in human serum.

Abstract

Alpha 1-acid glycoprotein (AGP) is an acute phase protein in mammals, including humans. The amount of AGP in human serum varies in response to certain diseases; thus, many efforts have been made to develop methods for quantifying human AGP. We recently discovered that luminescence occurs merely by mixing Cypridina luciferin with human AGP under human serum-free neutral or basic buffer conditions. In this study, we tested an application of Cypridina luciferin for quantifying AGP contained in human serum. Our luminescence spectrum measurements of Cypridina luciferin with human serum samples showed that the maximum emission wavelength with human serum (480nm) differed from that with human AGP (464nm) due to the abundant presence of endogenous human serum albumin (HSA). Furthermore, the luminescence intensities of Cypridina luciferin with human AGP in HSA-depleted human serum were consistent with those in a human serum-free basic buffer, but those in human serum were not. These results indicated that depletion of HSA in human serum was required to use Cypridina luciferin for quantifying AGP in human serum. Additionally, we found that the luminescence intensity of Cypridina luciferin with bovine AGP was approximately tenfold lower than that with human AGP.