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, non-specific 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.

中文翻译：

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用于验证血型试剂的RBC膜抗原阵列的开发

抗体试剂一直是临床中表征血型（BG）抗原的标准方法。这些BG抗体的特异性和交叉反应性是使用凝胶微柱测定（GMA）常规检测的。但是，GMA既不特异性也不敏感，因此增加了不正确匹配的RBC输血的风险。在这项工作中，我们描述了一种基于珠的RBC膜抗原阵列，可检测BG抗体-抗原结合，其灵敏度和动态范围比GMA高700倍。使用高度富集BG面板抗原的RBC碎片膜来制造RBC膜抗原阵列。然后将阵列用于筛选15种BG试剂与三个抗原板的相互作用。大多数抗体反应（即86.7％； 39/45）与使用GMA获得的反应一致。六个交叉反应，通过凝集抑制和基因分型分析证实了仅由我们的阵列鉴定的非特异性抗体反应（即13.3％； 6/45）。这些结果表明，我们的RBC膜抗原阵列在筛选BG抗体和提高RBC输血安全性方面具有巨大潜力。