Sensitive detection of idiotypic platelet-reactive alloantibodies by an electrical protein chip

• Published in: Biosensors & bioelectronics 2012-06, Vol.36 (1), p.207-211
• Main Authors: Quiel, Annett, Jürgen, Britta, Greinacher, Andreas, Lassen, Susan, Wörl, Ralf, Witt, Sabine, Schweder, Thomas
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal - chemistry; Antigens, Human Platelet - analysis; Biological and medical sciences; Biosensing Techniques; Biotechnology; Blood Platelet Disorders - diagnosis; Electrical protein chip; Electrochemical Techniques; Fundamental and applied biological sciences. Psychology; Gold; HLA Antigens - analysis; Human Platelet Antigen; Humans; Isoantibodies - analysis; Isoantibodies - blood; MAIPA; Microelectrodes; Protein Array Analysis - methods; Sandwich immunoassay; Sensitivity and Specificity; Thrombocytopenia; Thrombocytopenia - diagnosis
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2012.04.021

To prevent and treat immune-mediated platelet disorders (e.g. neonatal allo-immune thrombocytopenia and platelet transfusion refractoriness) the causative idiotypic platelet-reactive antibodies have to be detected with high sensitivity and specificity. The “Monoclonal Antibody Immobilization Platelet Assay” (MAIPA) is the diagnostic gold standard for immunotyping sera with respect to alloantibodies against human platelet antigens (HPA). However, it is labor-intensive and time-consuming. In this work, an automated protein chip assay (enzyme-linked sandwich immunoassay) based on interdigitated gold microelectrodes in combination with an electrical read-out system was developed and optimized. For this purpose, specific capture antibodies were immobilized on the gold electrodes. The binding of the target is detected via an enzyme-labeled detection antibody by a redox-recycling process that corresponds to the amount of bound target molecule. With this electrical chip assay it is possible to detect antibodies against HPA-1a, HPA-5b and HLA with high sensitivity and specificity in less than half the duration of the MAIPA protocol with similar intra- and interassay variance. ► An electrical assay for the detection of human alloantibodies was developed. ► The protein chip enables a fast detection of idiotypic platelet-reactive antibodies. ► The detection is based on an automated electrical enzyme-linked sandwich immunoassay. ► An averaged intra- and inter-assay variability below 10% was reached.