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通过细胞表面等离子体共振成像进行多重血型分型。

Multiplex blood group typing by cellular surface plasmon resonance imaging.

作者信息

Szittner Zoltan, Bentlage Arthur E H, van der Donk Eric, Ligthart Peter C, Lissenberg-Thunnissen Suzanne, van der Schoot C Ellen, Vidarsson Gestur

机构信息

Department of Experimental Immunohematology, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

Department of Reagents, Sanquin Research and Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands.

出版信息

Transfusion. 2019 Feb;59(2):754-761. doi: 10.1111/trf.15071. Epub 2018 Nov 29.

Abstract

BACKGROUND

Blood-group typing of donors and patients is essential to avoid incompatible transfusions. Transfusion of incompatible RBCs may result in alloimmunization complicating future transfusions or in the presence of antibodies in adverse reactions. With more than 300 blood group antigens identified, it is difficult to provide fully compatible blood. Currently, standard practice is to match for the most immunogenic antigens. While the current agglutination-based RBC-typing methods are reliable for testing a selected number of antigens, they are not easily adaptable for high-throughput multiplex blood typing beyond the current standard.

STUDY DESIGN AND METHODS

Surface plasmon resonance (SPR) is a label-free method to follow molecular-and, very recently, also cellular-interactions in real time. Demonstration of binding of RBCs to blood group antigen-specific antibodies by SPR has already been achieved. Here, we demonstrate the generation of an SPR array equipped with clinically relevant blood group antibodies (A, B, and Rh blood groups). To validate this method, we blindly compared typing of 946 blood donors with results of current diagnostic agglutination-based methods.

RESULTS

RBC typing was achieved by monitoring RBC binding to blood group-specific antibodies on the sensor simultaneously within 5 minutes per sample. Regeneration of the chip was robust, allowing for typing of at least 100 samples. The typing results gave a 100% match with classical serology with all antibodies tested besides anti-E/e monoclonals, which gave inconsistent results due to low antibody specificity.

CONCLUSION

This study demonstrates that SPR-based RBC typing for multiple antigens can be realized simultaneously with high-quality antibodies, enabling reduced hands-on time and possibly improving cost efficiency.

摘要

背景

对献血者和患者进行血型鉴定对于避免不相容输血至关重要。输注不相容的红细胞可能导致同种免疫,使未来输血变得复杂,或在不良反应中出现抗体。已鉴定出300多种血型抗原,提供完全相容的血液很困难。目前的标准做法是匹配最具免疫原性的抗原。虽然当前基于凝集的红细胞血型鉴定方法对于检测选定数量的抗原是可靠的,但它们不容易适应超出当前标准的高通量多重血型鉴定。

研究设计与方法

表面等离子体共振(SPR)是一种无需标记的方法,可实时跟踪分子以及最近的细胞相互作用。通过SPR已实现红细胞与血型抗原特异性抗体结合的证明。在此,我们展示了配备临床相关血型抗体(A、B和Rh血型)的SPR阵列的生成。为了验证该方法,我们将946名献血者的血型鉴定结果与当前基于凝集的诊断方法的结果进行了盲法比较。

结果

通过监测每个样本在5分钟内红细胞与传感器上血型特异性抗体的结合来实现红细胞血型鉴定。芯片的再生性能良好,允许对至少100个样本进行血型鉴定。除抗E/e单克隆抗体外,所有测试抗体的血型鉴定结果与经典血清学结果100%匹配,抗E/e单克隆抗体由于抗体特异性低而给出不一致的结果。

结论

本研究表明,基于SPR的多种抗原红细胞血型鉴定可以与高质量抗体同时实现,从而减少实际操作时间并可能提高成本效益。

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