Development of a method for measuring complement C1s protein on platelet surface and its preliminary application
10.13602/j.cnki.jcls.2025.11.06
- VernacularTitle:血小板表面C1s蛋白表达检测方法的建立及初步应用
- Author:
Jun YE
1
;
Huimin LU
;
Jianfeng ZHU
;
Huilian HUA
;
Xin XU
;
Yili YANG
;
Chao MENG
;
Min SHA
Author Information
1. 泰州市人民医院中心实验室,江苏泰州 225300
- Publication Type:Journal Article
- Keywords:
complement C1s;
magnetic beads,platelet;
idiopathic thrombocytopenic purpura;
flow cytometry
- From:
Chinese Journal of Clinical Laboratory Science
2025;43(11):830-835
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a flow cytometry method for detecting C1s protein on platelet surface and preliminarily explore its potential application value in the auxiliary diagnosis of primary immune thrombocytopenia(ITP).Methods C1s-conjugated 2 μm car-boxylated magnetic beads(C1s beads)were prepared and used as quality control particles.Fluorescein isothiocyanate(FITC)-labeled anti-C1s antibody was employed as the detection antibody to develop a flow cytometric assay for detecting C1s protein expression on platelets.The intra-assay and inter-assay precision,as well as the dilution linearity of the method,were evaluated.Subsequently,the expression levels of C1 s protein on the surface of platelets were compared among the ITP group,the non-ITP thrombocytopenia group,and the healthy control group.Results Light microscopy showed that both unconjugated carboxylated magnetic beads(blank beads)and C1s-conjugated beads were uniformly dispersed without aggregation.Under fluorescence microscopy,C1s beads exhibited strong yellow-green fluorescence,whereas the blank beads showed no fluorescence signal.The established flow cytometry assay exhibited ac-ceptable precision,with intra-assay coefficient of variation(CV)values of 7.02％,7.12％,and 3.91％for low,medium,and high con-centrations of C1s beads,respectively,and inter-assay CV values of 13.49％,6.15％,and 0.78％,respectively.The dilution linearity was satisfactory,coefficient of determination(R2)=0.998 8.Clinical sample testing revealed that the proportion of C1s-positive plate-lets in ITP group(2.56±0.79)％was significantly higher than that in healthy control group(0.23±0.18)％and the non-ITP thrombo-cytopenia control group(0.22±0.10)％,with statistically significant differences(both P＜0.05).Conclusion This study successfully established a stable and reliable flow-cytometry method for quantifying C1s expression on platelet surface and preliminarily demonstrated that C1s expression is significantly elevated on platelets of ITP patients,suggesting that C1s could serve as a potential auxiliary diag-nostic marker for ITP.
