title: High-throughput allogeneic antibody detection using protein microarrays.
abstract: 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.
authors.affiliation: 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.
authors.forename: Jed
authors.initials: J
authors.lastname: Paul
authors.affiliation: 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.
authors.forename: Bita
authors.initials: B
authors.lastname: Sahaf
authors.affiliation: 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.
authors.forename: Spenser
authors.initials: S
authors.lastname: Perloff
authors.affiliation: Division of Blood and Marrow Transplantation, Stanford University School of Medicine, 269 West Campus Dr., CCSR #2205, Stanford, CA 94305, United States.
authors.forename: Kelsi
authors.initials: K
authors.lastname: Schoenrock
authors.affiliation: 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.
authors.forename: Fang
authors.initials: F
authors.lastname: Wu
authors.affiliation: 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.
authors.forename: Hideki
authors.initials: H
authors.lastname: Nakasone
authors.affiliation: 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.
authors.forename: John
authors.initials: J
authors.lastname: Coller
authors.affiliation: 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.
authors.forename: David
authors.initials: D
authors.lastname: Miklos
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000097:blood; D002648:Child; D002908:Chronic Disease; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D006086:Graft vs Host Disease / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D006182:H-Y Antigen / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D057166:High-Throughput Screening Assays; D006648:Histocompatibility; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007518:Isoantibodies / Q000097:blood* / Q000276:immunology; D008297:Male; D008875:Middle Aged; D011237:Predictive Value of Tests; D040081:Protein Array Analysis / Q000379:methods*; D015203:Reproducibility of Results; D055815:Young Adult
chemical_list: D015415:Biomarkers; D000939:Epitopes; D006182:H-Y Antigen; D007518:Isoantibodies
keywords: Alloantibody; Chronic graft versus host disease; HY antigen; Hematopoietic cell transplantation; Protein microarray