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使用基于昆虫细胞的系统鉴定和定量人血清中的抗Gp.Mur抗体

Identification and Quantification of Anti-Gp.Mur Antibodies in Human Serum Using an Insect-Cell-Based System.

作者信息

Lamis Robert John S, Chiueh Tsong-Shi, Tsai Chih-Hsuan, Lo Huei-Ru, Wei Sung-Chan, Chao Yu-Chan

机构信息

Molecular and Cell Biology, National Defense Medical Center, Taiwan International Graduate Program, Academia Sinica and Graduate Institute of Life Science, Taipei 115, Taiwan.

Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan.

出版信息

Diagnostics (Basel). 2021 May 27;11(6):966. doi: 10.3390/diagnostics11060966.

Abstract

Gp.Mur is a clinically relevant antigen of the MNS blood group system that is highly prevalent in several Asian populations. Its corresponding antibody, anti-Gp.Mur, has been implicated in hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. Currently, identifying and confirming anti-Gp.Mur antibody presence in sera via agglutination of a panel of red blood cells (RBCs) is inefficient and difficult to quantify. Using a baculovirus expression system to express Gp.Mur antigen on insect cell surfaces, we have developed a quantitative cell-based system to confirm the presence of anti-Gp.Mur antibody in human serum. We obtained 10 serum samples preidentified as having anti-Gp.Mur antibody and another 4 samples containing noncorresponding antibodies from hospital patients. Insect cells displaying Gp.Mur antigen successfully adsorbed anti-Gp.Mur antibody in the sera and inhibited the RBC agglutination mediated by this antibody. By varying the concentration of Gp.Mur-displaying cells, we could grade levels of RBC agglutination by anti-Gp.Mur antibody. Densitometric analysis further enabled quantitative determinations of hemagglutination inhibition by Gp.Mur-displaying cells. We believe that this cell-based hemagglutination inhibition system greatly improves or supplements existing technology and is a convenient means for accurately identifying and quantifying anti-Gp.Mur antibody.

摘要

Gp.Mur是MNS血型系统中一种具有临床相关性的抗原,在多个亚洲人群中高度流行。其相应抗体抗-Gp.Mur与溶血性输血反应以及胎儿和新生儿溶血病有关。目前,通过一组红细胞(RBC)的凝集来鉴定和确认血清中抗-Gp.Mur抗体的存在效率低下且难以量化。利用杆状病毒表达系统在昆虫细胞表面表达Gp.Mur抗原,我们开发了一种基于细胞的定量系统来确认人血清中抗-Gp.Mur抗体的存在。我们从医院患者中获得了10份预先鉴定为含有抗-Gp.Mur抗体的血清样本以及另外4份含有非相应抗体的样本。展示Gp.Mur抗原的昆虫细胞成功吸附了血清中的抗-Gp.Mur抗体,并抑制了该抗体介导的RBC凝集。通过改变展示Gp.Mur的细胞浓度,我们可以对抗-Gp.Mur抗体引起的RBC凝集程度进行分级。光密度分析进一步能够对展示Gp.Mur的细胞介导的血凝抑制进行定量测定。我们认为这种基于细胞的血凝抑制系统极大地改进或补充了现有技术,是准确鉴定和定量抗-Gp.Mur抗体的便捷方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aefd/8227843/7622327c8392/diagnostics-11-00966-g001.jpg

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