High-throughput allogeneic antibody detection using protein microarrays

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• 作者：Jed Paul^a ; ^{jedpaul@stanford.edu" class="auth_mail" title="E-mail the corresponding author} ; Bita Sahaf^a ; ^{bsahaf@stanford.edu" class="auth_mail" title="E-mail the corresponding author} ; Spenser Perloff^a ; ^{shperloff@ucdavis.edu" class="auth_mail" title="E-mail the corresponding author} ; Kelsi Schoenrock^a ; Fang Wu^a ; ^{fangw@stanford.edu" class="auth_mail" title="E-mail the corresponding author} ; Hideki Nakasone^a ; ^{nakasone-tky@umin.ac.jp" class="auth_mail" title="E-mail the corresponding author} ; John Coller^b ; ^{john.coller@stanford.edu" class="auth_mail" title="E-mail the corresponding author} ; David Miklos^a ; ^{dmiklos@stanford.edu" class="auth_mail" title="E-mail the corresponding author} ; ^{dmiklos@stanord.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：ELISA ; enzyme-linked immunosorbent assay ; HCT ; hematopoietic cell transplantation ; mHA ; minor histocompatibility antigen ; cGVHD ; chronic graft versus host disease ; F➔M ; female donor into a male recipient ; MFI ; median fluorescence intensity ; IQR ; interquartile range
• 刊名：Journal of Immunological Methods
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：432
• 期：Complete
• 页码：57-64
• 全文大小：1176 K

文摘

Enzyme-linked immunosorbent assays (ELISAs) have traditionally been used to detect alloantibodies in patient plasma samples post hematopoietic cell transplantation (HCT); however, protein microarrays have the potential to be multiplexed, more sensitive, and higher throughput than ELISAs. Here, we describe the development of a novel and sensitive microarray method for detection of allogeneic antibodies against minor histocompatibility antigens encoded on the Y chromosome, called HY antigens. Six microarray surfaces were tested for their ability to bind recombinant protein and peptide HY antigens. Significant allogeneic immune responses were determined in male patients with female donors by considering normal male donor responses as baseline. HY microarray results were also compared with our previous ELISA results. Our overall goal was to maximize antibody detection for both recombinant protein and peptide epitopes. For detection of HY antigens, the Epoxy (Schott) protein microarray surface was both most sensitive and reliable and has become the standard surface in our microarray platform.