Chen Qingqing, Li Fei, Wang Haoran, Bu Changkai, Shi Feng, Jin Lan, Zhang Qunye, Chi Lianli
National Glycoengineering Research Center, Shandong University, Qingdao, China.
Scientific Research Division, Shandong Institute for Food and Drug Control, Jinan, China.
Front Mol Biosci. 2022 Aug 26;9:966754. doi: 10.3389/fmolb.2022.966754. eCollection 2022.
Heparin (HP) is a polysaccharide that is widely used in the clinic as an anticoagulant. A major side effect associated with HP is the heparin-induced thrombocytopenia (HIT), which is initiated by the immune response to complex formed by HP and platelet factor 4 (PF4). Low molecular weight heparins (LMWHs) are the depolymerized version of HP, which have reduced risks of inducing HIT. However, it is still necessary to evaluate the immunogenicity of LMWHs to ensure their drug safety. Since HIT involves very complicated processes, the evaluation of HP and LMWH immunogenicity requires experiments from multiple aspects, of which the binding affinity between HP and PF4 is a key property to be monitored. Herein, we developed a novel competitive biolayer interferometry (BLI) method to investigate the binding affinity between HP and PF4. The influence of different domains in HP on its immunogenicity was compared for better understanding of the molecular mechanism of HP immunogenicity. Furthermore, the half maximal inhibitory concentration (IC) of HP and LMWH can be measured by competitive combination, which is important for the quality control during the developing and manufacturing of HP and LMWH drugs.
肝素（HP）是一种多糖，在临床上广泛用作抗凝剂。与HP相关的一个主要副作用是肝素诱导的血小板减少症（HIT），它是由对HP与血小板因子4（PF4）形成的复合物的免疫反应引发的。低分子量肝素（LMWHs）是HP的解聚形式，其诱导HIT的风险较低。然而，评估LMWHs的免疫原性以确保其药物安全性仍然很有必要。由于HIT涉及非常复杂的过程，对HP和LMWH免疫原性的评估需要从多个方面进行实验，其中HP与PF4之间的结合亲和力是一个需要监测的关键特性。在此，我们开发了一种新型的竞争性生物层干涉术（BLI）方法来研究HP与PF4之间的结合亲和力。比较了HP中不同结构域对其免疫原性的影响，以更好地理解HP免疫原性的分子机制。此外，HP和LMWH的半数最大抑制浓度（IC）可以通过竞争性结合来测量，这对于HP和LMWH药物研发和生产过程中的质量控制很重要。
