TY - JOUR
T1 - Development of a recombinant anti-Vel immunoglobulin M to identify Vel-negative donors
AU - van der Rijst, Marea V. E.
AU - Lissenberg-Thunnissen, Suzanne N.
AU - Ligthart, Peter C.
AU - Visser, Remco
AU - Jongerius, John M.
AU - Voorn, Lesley
AU - Veldhuisen, Barbera
AU - Vidarsson, Gestur
AU - van den Akker, Emile
AU - van der Schoot, C. Ellen
N1 - Funding Information: From the 1Department of Hematopoiesis, Sanquin Research and Landsteiner Laboratory, AUMC, Amsterdam, The Netherlands; 2Department of Experimental Immunohematology, Sanquin Research and Landsteiner Laboratory, AUMC, Amsterdam, The Netherlands; 3Department of Immunohematology Diagnostic Services, Sanquin, Amsterdam, The Netherlands; and the 4Department of Research and Lab Services, National Screening Laboratory Sanquin, Sanquin, Amsterdam, the Netherlands. Address reprint requests to: C. Ellen van der Schoot, PhD, Ples-manlaan 125, 1066 CX, Amsterdam, the Netherlands; e-mail: e. vanderschoot@sanquin.nl. This work was supported by grants from Landsteiner Foundation (LSBR: 1351; MVEvdR, EvdA, CEvdS) and Sanquin (PPODR: 14-13; BV). Received for publication September 20, 2018; revision received November 28, 2018, and accepted December 3, 2018. doi:10.1111/trf.15147 © 2019 AABB TRANSFUSION 2019;59;1359–1366 Publisher Copyright: © 2019 AABB
PY - 2019
Y1 - 2019
N2 - BACKGROUND: Alloimmunization against the high-frequency Vel blood group antigen may result in transfusion reactions or hemolytic disease of fetus and newborn. Patients with anti-Vel alloantibodies require Vel-negative blood but Vel-negative individuals are rare (1:4000). Identification of Vel-negative donors ensures availability of Vel-negative blood; however, accurate Vel blood group typing is difficult due to variable Vel antigen expression and limited availability of anti-Vel typing sera. We report the production of a recombinant anti-Vel that also identifies weak Vel expression. STUDY DESIGN AND METHODS: A recombinant anti-Vel monoclonal antibody was produced by cloning the variable regions from an anti-Vel–specific B cell isolated from an alloimmunized patient into a vector harboring the constant regions of immunoglobulin (Ig)G1-kappa or IgM-kappa. Antibody Vel specificity was tested by reactivity to SMIM1-transfected HEK293T cells and by testing various red blood cells (RBCs) of donors with normal, weak, or no Vel expression. High-throughput donor screening applicability was tested using an automated blood group analyzer. RESULTS: A Vel-specific IgM class antibody was produced. The antibody was able to distinguish between Vel-negative and very weak Vel antigen–expressing RBCs by direct agglutination and in high-throughput settings using a fully automated blood group analyzer and performed better than currently used human anti-Vel sera. High-throughput screening of 13,288 blood donations identified three new Vel-negative donors. CONCLUSION: We generated a directly agglutinating recombinant anti-Vel IgM, M3F5S-IgM, functional in manual, automated agglutination assays and flow cytometry settings. This IgM anti-Vel will improve diagnostics by facilitating the identification of Vel-negative blood donors.
AB - BACKGROUND: Alloimmunization against the high-frequency Vel blood group antigen may result in transfusion reactions or hemolytic disease of fetus and newborn. Patients with anti-Vel alloantibodies require Vel-negative blood but Vel-negative individuals are rare (1:4000). Identification of Vel-negative donors ensures availability of Vel-negative blood; however, accurate Vel blood group typing is difficult due to variable Vel antigen expression and limited availability of anti-Vel typing sera. We report the production of a recombinant anti-Vel that also identifies weak Vel expression. STUDY DESIGN AND METHODS: A recombinant anti-Vel monoclonal antibody was produced by cloning the variable regions from an anti-Vel–specific B cell isolated from an alloimmunized patient into a vector harboring the constant regions of immunoglobulin (Ig)G1-kappa or IgM-kappa. Antibody Vel specificity was tested by reactivity to SMIM1-transfected HEK293T cells and by testing various red blood cells (RBCs) of donors with normal, weak, or no Vel expression. High-throughput donor screening applicability was tested using an automated blood group analyzer. RESULTS: A Vel-specific IgM class antibody was produced. The antibody was able to distinguish between Vel-negative and very weak Vel antigen–expressing RBCs by direct agglutination and in high-throughput settings using a fully automated blood group analyzer and performed better than currently used human anti-Vel sera. High-throughput screening of 13,288 blood donations identified three new Vel-negative donors. CONCLUSION: We generated a directly agglutinating recombinant anti-Vel IgM, M3F5S-IgM, functional in manual, automated agglutination assays and flow cytometry settings. This IgM anti-Vel will improve diagnostics by facilitating the identification of Vel-negative blood donors.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85060862633&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/30702752
U2 - https://doi.org/10.1111/trf.15147
DO - https://doi.org/10.1111/trf.15147
M3 - Article
C2 - 30702752
SN - 0041-1132
VL - 59
SP - 1359
EP - 1366
JO - Transfusion
JF - Transfusion
IS - 4
ER -