Document Type : Original Article
Authors
1
Immunogenetics Research Center, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
2
Department of Hematology and Medical Laboratory Sciences, Faculty of Allied Medicine, Kerman University of Medical Sciences, Kerman, Iran
3
Cell Therapy and Hematopoietic Stem Cell Transplantation Research Center, Research Institute for Oncology, Hematology and Cell Therapy, Tehran University of Medical Sciences, Tehran, Iran
4
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
10.22034/iji.2026.112881.3294
Abstract
Background: Antiplatelet antibodies may develop following platelet transfusion, during pregnancy, and in autoimmune diseases such as immune thrombocytopenic purpura. This substantiates the use of a proper screening method for diagnosis and clinical management.
Objectives: This study focuses on the design and production of an ELISA-based screening kit for antiplatelet antibodies.
Methods: Serum and platelet samples were obtained from patients with platelet refractoriness and healthy donors. Three experimental approaches were applied to eliminate nonspecific binding: removing IgG bound to platelet FcγRIIA, preventing the rebinding of serum IgG to FcγRIIA, and inhibiting the secondary antibody from detecting IgG attached to FcγRIIA.
Results: Physical and chemical methods removed IgG bound to FcγRIIA, but they could not prevent FcγRIIA rebinding after serum addition. Also, pre-treatment of sera with rabbit polyclonal anti-Fc antibodies effectively prevented FcγRIIA rebinding. Moreover, using a secondary antibody against the CH2 domain could distinguish antibodies bound to platelet antigens from those bound to FcγRIIA.
Conclusion: The application of a monoclonal antibody targeting the CH2 domain represents an optimal method for evaluating IgG associated with intact platelet antigens, eliminating the need for supplementary treatment procedures.
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