Scandinavian Journal of Clinical and Laboratory Investigation ( IF 2.1 ) Pub Date : 2022-09-21 , DOI: 10.1080/00365513.2022.2122077 Maria Kløjgaard Skytthe 1 , Anna Lahn Sørensen 1 , Dorle Hennig 1 , Maria Boysen Sandberg 2 , Lars Melholt Rasmussen 2 , Holger J Møller 3, 4 , Karsten Skjødt 1 , Jonas Heilskov Graversen 1 , Søren Kragh Moestrup 1, 4, 5
Abstract
Haptoglobin (Hp) is an abundant plasma protein scavenging hemoglobin (Hb) via CD163 on macrophages. This process consumes Hp, which therefore negatively correlates to hemolysis. However, exact measurements of Hp plasma levels are complicated by different phenotypes (Hp1-1, Hp2-1, and Hp2-2) forming different oligomeric states with differences in immunoreactivity. In addition, humans have an immune-cross-reactive Hp-related protein. In the present study, we developed Hp-specific monoclonal antibodies for an accurate Hp analysis of the different Hp phenotypes in a panel of 112 anonymous samples from hospitalized individuals subjected to routine Hp immunoturbidimetric measurements. The data revealed immunoturbidimetry as a reliable method in most cases but also that the use of non-phenotype-specific calibrators leads to substantial bias in the measurement of the Hp-concentration, non at least in Hp1-1 individuals. Furthermore, analysis of the Hb-dependence of the CD163 interaction with Hp1-1 and Hp2-2 showed that a higher ‘cost-effectiveness’ in the consumption of dimeric Hp1-1 versus multimeric Hp phenotypes is a likely contribution to the observed differences in the plasma levels of the Hp phenotypes. In conclusion, the determination of Hp phenotype and the use of phenotype-specific calibrators are essential to obtain a precise estimate of the Hp level in healthy and diseased individuals.
中文翻译:
重新评估人血浆样品中触珠蛋白的测量
摘要
触珠蛋白 (Hp) 是一种丰富的血浆蛋白,可通过以下途径清除血红蛋白 (Hb)巨噬细胞上的 CD163。该过程消耗 Hp,因此与溶血呈负相关。然而,由于不同的表型(Hp1-1、Hp2-1 和 Hp2-2)形成具有不同免疫反应性的不同寡聚状态,因此 Hp 血浆水平的精确测量变得复杂。此外,人类具有免疫交叉反应性 Hp 相关蛋白。在本研究中,我们开发了 Hp 特异性单克隆抗体,用于对来自接受常规 Hp 免疫比浊测量的住院患者的 112 个匿名样本的不同 Hp 表型进行准确的 Hp 分析。数据显示,在大多数情况下,免疫比浊法是一种可靠的方法,但使用非表型特异性校准物会导致 Hp 浓度测量存在重大偏差,至少在 Hp1-1 个体中是这样。此外,对 CD163 与 Hp1-1 和 Hp2-2 相互作用的 Hb 依赖性的分析表明,与多聚体 Hp 表型相比,二聚体 Hp1-1 的消耗具有更高的“成本效益”,这可能有助于观察到血浆中的差异Hp 表型水平。总之,Hp 表型的测定和表型特异性校准器的使用对于准确估计健康和患病个体的 Hp 水平至关重要。