Quantitative detection of red blood cell antibody-mediated complement activation
Journal Title: Chinese Journal of Blood Transfusion - Year 2022, Vol 35, Issue 9
Abstract
Objective To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. Methods Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. Results The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tja 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tja mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. Conclusion In vitro model of anti-Tja-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.
Authors and Affiliations
Zhongying WANG, Jian LI, Fengyong ZHAO, Chenrui QIAN, Wei SHEN, Liangfeng FAN, Sha JIN, Jiewei ZHENG, Yuyu ZHANG, Dong XIANG
Keywords
Related Articles
- EP ID EP738359
- DOI 10.13303/j.cjbt.issn.1004-549x.2022.09.025
- Views 6
- Downloads 0
Bibliometrics and visual analysis of platelet rich plasma based on strategic coordinate diagram and co-word analysis
Objective To explore the research status, hot spots and trends of platelet rich plasma (PRP). Methods With "platelet rich plasma (PRP)" and its Chinese equivalent as the subject words, the PRP related articles during Jan...
Preliminary discussion on the causes and influencing factors of blood discarding
Objective To conduct a retrospective statistical analysis of the blood discarding situation in Xuzhou Central Blood Station in the past 5 years, so as to explore relevant measures and strategies to reduce blood discardin...
A survey on professional identity and job satisfaction of blood transfusion physicians and its influencing factors, in Tianjin
Objective To investigate the status of professional identity and job satisfaction of blood transfusion physicians in clinical in Tianjin, and to analyze the influencing factors. Methods The status of professional identit...
Application and evaluation of human antibodies in ABO grouping
Objective To evaluate the application of monoclonal typing reagents and human anti-A/B antibodies for absorption-elution test in ABO grouping. Methods The specificity of monoclonal typing reagents and human anti-A/B anti...
Effect of X-ray irradiation on the inactivation of lymphocytes and the quality of apheresis platelets
Objective To investigate the inactivation function of 25Gy X-ray irradiation on apheresis platelets’ lymphocytes and its effect on the quantity of apheresis platelets(AP). Methods Twenty healthy voluntary AP donors from...