Development of RBC Membrane Antigen Arrays for Validating Blood Grouping Reagents
Development of RBC Membrane Antigen Arrays for Validating Blood Grouping Reagents
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开发用于验证血型试剂的红细胞膜抗原阵列
DOI：
10.1021/acs.jproteome.8b00370
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Xiaobo Yu
中科院分区：
文献类型：
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作者：
Lu Yang;Xiaobo Yu
抗体试剂在临床上一直是鉴定血型（BG）抗原的标准方法。这些血型抗体的特异性和交叉反应性通常使用凝胶微柱法（GMA）进行检测。然而，凝胶微柱法既不特异也不灵敏，从而增加了红细胞输注不匹配的风险。在这项工作中，我们描述了一种基于微珠的红细胞膜抗原阵列，其检测血型抗体 - 抗原结合的灵敏度和动态范围比凝胶微柱法高约700倍。红细胞膜抗原阵列是使用高度富集血型抗原组的破碎红细胞膜制备的。然后使用这些阵列筛选15种血型试剂与三种抗原组的相互作用。大多数抗体反应（即86.7%；45个中的39个）与凝胶微柱法获得的结果一致。仅由我们的阵列鉴定出的6种交叉反应、非特异性抗体反应（即13.3%；45个中的6个）通过凝集抑制和基因分型试验得到了证实。这些结果表明，我们的红细胞膜抗原阵列在筛选血型抗体和提高红细胞输注安全性方面具有巨大潜力。
Antibody reagents have been remained as a standard approach to characterize blood group (BG) antigens in clinic. The specificity and cross-reactivity of these BG antibodies are routine detected using the gel microcolumn assay (GMA). However, the GMA is neither specific nor sensitive, thus increasing the risk of improperly matched RBC transfusions. In this work, we describe a bead-based RBC membrane antigen array to detect BG antibody-antigen binding with ∼700-fold higher sensitivity and dynamic range than the GMA. RBC membrane antigen arrays were fabricated using fragmented RBC membranes highly enriched in BG panel antigens. The arrays were then used to screen the interactions of 15 BG reagents to three antigen panels. The majority of the antibody reactions (i.e., 86.7%; 39/45) aligned with those obtained with the GMA. The six cross-reactive, nonspecific antibody reactions identified only by our arrays (i.e., 13.3%; 6/45) were confirmed by agglutination inhibition and genotyping assays. These results demonstrate that our RBC membrane antigen array has great potential in screening BG antibodies and improving the safety of RBC transfusions.