[Establishment of Quantitative SPR Assay for Antibodies Against Human Platelet Antigen-1a].

Zhongguo Shi Yan Xue Ye Xue Za Zhi

Shanghai Blood Center, Shanghai Institute of Blood Transfusion, Shanghai 200051, China,E-mail:

Published: February 2021

Objective: To establish quantitative surface plasmon resonance (SPR) assay for antibodies against human platelet antigen-1a (HPA-1a).

Methods: Recombinant protein was fixed on the chip surface by amino coupling method. SPR assay was used to detect the standard antibodies against HPA-1a at different conceatration. The optimal experimental parameters were determined, and standard curves were constructed with linear regression. Moreover, the sensitivity, specificity, accuracy and precision of the assay were evaluated.

Results: The quantitative SPR assay for HPA-1a antibodies was established. The determination ranges were 0-20 IU, with accuracy (recovery rate) was 97.75%-103.08%. The intra-assay precision [coefficients of variation (CV)] was 3.53%-4.29%, and the inter-assay precision (CV) was 2.08%-4.40%. For specificity test, several kinds of monoclonal and human antibodies against platelet membrane protein were tested and no positive result was observed.

Conclusion: The established quantitative SPR assay for HPA-1a antibodies shows good sensitivity, specificity, accuracy and precision, and this rapid and simple method provides a new reference method for scientific research and clinical antibody detection.

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Source
http://dx.doi.org/10.19746/j.cnki.issn.1009-2137.2021.01.039DOI Listing

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