A rapid one‐stage whole‐blood HPA‐1a phenotyping assay using a recombinant monoclonal IgG1 anti‐HPA‐1a

Severe neonatal alloimmune thrombocytopenia and patients with HPA‐1a‐specific antibodies require transfusion of HPA‐1a‐negative platelets. Identifying HPA‐1a‐negative donors requires simple and reliable typing methods. Most existing techniques use polyclonal antibodies, are time consuming and involv...

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Bibliographic Details
Published in:British journal of haematology 2000-02, Vol.108 (2), p.440-447
Main Authors: GARNER, S. F, SMETHURST, P. A, CLARK, M. R, RIGAL, D, SCHAWALLER, M, OUWEHAND, W. H, MERIEUX, Y, AEBY, C, SMITH, G, ARMOUR, K. L, SCOTT, M. L, WILLIAMSON, L. M, METCALFE, P, GOODALL, A. H
Format: Article
Language:English
Subjects:
Antigens, Human Platelet - genetics
Biological and medical sciences
Blood Grouping and Crossmatching - methods
Enzyme-Linked Immunosorbent Assay - methods
enzyme‐linked immunosorbent assay
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Genotype
Hematology
HPA‐1a typing
Humans
Immunoglobulin G - analysis
Immunohematology
Phenotype
Platelet immunology
platelets
recombinant monoclonal anti‐HPA‐1a
whole blood
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Summary:Severe neonatal alloimmune thrombocytopenia and patients with HPA‐1a‐specific antibodies require transfusion of HPA‐1a‐negative platelets. Identifying HPA‐1a‐negative donors requires simple and reliable typing methods. Most existing techniques use polyclonal antibodies, are time consuming and involve platelet isolation. We have used a horseradish peroxidase (HRP)‐conjugated recombinant IgG1 anti‐HPA‐1a (CAMTRAN007) to develop a rapid and reliable enzyme‐linked immunosorbent assay (ELISA), which eliminates sample preparation and reduces the incubation and wash steps associated with traditional sandwich ELISAs. The assay uses simultaneous incubation of the monoclonal antibody RFGP56 to capture GPIIbIIIa from whole blood and the recombinant IgG1 antibody to detect captured HPA‐1a antigen. It allows 96 samples to be typed in less than 1 h and can be used on stored samples. Initial testing of 85 samples of known HPA‐1a genotype demonstrated that HPA‐1a‐negative samples had OD values of  0.6. Testing of 1862 random donor samples in two blood centres confirmed these OD cut‐off values and identified 45 HPA‐1a‐negative samples (2.4%), all except one giving OD values of
ISSN:0007-1048
1365-2141
DOI:10.1046/j.1365-2141.2000.01839.x