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High-throughput allogeneic antibody detection using protein microarrays.
Paul, Jed; Sahaf, Bita; Perloff, Spenser; Schoenrock, Kelsi; Wu, Fang; Nakasone, Hideki; Coller, John; Miklos, David.
Afiliação
  • Paul J; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States. Electronic address: jedpaul@stanford.edu.
  • Sahaf B; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States. Electronic address: bsahaf@stanford.edu.
  • Perloff S; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States. Electronic address: shperloff@ucdavis.edu.
  • Schoenrock K; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States.
  • Wu F; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States. Electronic address: fangw@stanford.edu.
  • Nakasone H; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States. Electronic address: nakasone-tky@umin.ac.jp.
  • Coller J; Stanford Functional Genomics Facility, Stanford University School of Medicine, 269 West Campus Dr., CCSR #0120, Stanford, CA 94305, United States. Electronic address: john.coller@stanford.edu.
  • Miklos D; Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States. Electronic address: dmiklos@stanford.edu.
J Immunol Methods ; 432: 57-64, 2016 May.
Article em En | MEDLINE | ID: mdl-26902899
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Teste de Histocompatibilidade / Antígeno H-Y / Transplante de Células-Tronco Hematopoéticas / Análise Serial de Proteínas / Ensaios de Triagem em Larga Escala / Doença Enxerto-Hospedeiro / Histocompatibilidade / Isoanticorpos Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Adolescent / Adult / Aged / Child / Female / Humans / Male / Middle aged Idioma: En Ano de publicação: 2016 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Teste de Histocompatibilidade / Antígeno H-Y / Transplante de Células-Tronco Hematopoéticas / Análise Serial de Proteínas / Ensaios de Triagem em Larga Escala / Doença Enxerto-Hospedeiro / Histocompatibilidade / Isoanticorpos Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Adolescent / Adult / Aged / Child / Female / Humans / Male / Middle aged Idioma: En Ano de publicação: 2016 Tipo de documento: Article