Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Hokkaido Center, Sapporo, 062-8517, Japan.
Anal Sci. 2022 Dec;38(12):1555-1562. doi: 10.1007/s44211-022-00191-7. Epub 2022 Oct 7.
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 (480 nm) differed from that with human AGP (464 nm) 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.
α 1-酸性糖蛋白(AGP)是哺乳动物(包括人类)的一种急性期蛋白。人血清中的 AGP 量会响应某些疾病而发生变化;因此,人们已经做出了许多努力来开发定量人 AGP 的方法。我们最近发现,在无人类血清的中性或碱性缓冲条件下,只需将海肾萤光素与人 AGP 混合,就会发生发光现象。在这项研究中,我们测试了海肾萤光素在定量人血清中 AGP 中的应用。我们对人血清样本中海肾萤光素的发光光谱测量表明,由于内源性人血清白蛋白(HSA)的大量存在,与人 AGP(464nm)相比,人血清中的最大发射波长(480nm)有所不同。此外,在 HSA 耗尽的人血清中,与人 AGP 混合的海肾萤光素的发光强度与人血清-free 碱性缓冲液中的发光强度一致,但在人血清中则不一致。这些结果表明,需要耗尽人血清中的 HSA,才能使用海肾萤光素定量人血清中的 AGP。此外,我们发现与人 AGP 相比,牛 AGP 与人海肾萤光素的发光强度大约低十倍。