Sensitive detection of idiotypic platelet-reactive alloantibodies by an electrical protein chip
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Sensitive detection of idiotypic platelet-reactive alloantibodies by an electrical protein chip. / Quiel, Annett; Jürgen, Britta; Greinacher, Andreas; Lassen, Susan; Wörl, Ralf; Witt, Sabine; Schweder, Thomas.
in: BIOSENS BIOELECTRON, Jahrgang 36, Nr. 1, 11.05.2012, S. 207-11.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Sensitive detection of idiotypic platelet-reactive alloantibodies by an electrical protein chip
AU - Quiel, Annett
AU - Jürgen, Britta
AU - Greinacher, Andreas
AU - Lassen, Susan
AU - Wörl, Ralf
AU - Witt, Sabine
AU - Schweder, Thomas
N1 - Copyright © 2012 Elsevier B.V. All rights reserved.
PY - 2012/5/11
Y1 - 2012/5/11
N2 - 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.
AB - 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.
KW - Antibodies, Monoclonal/chemistry
KW - Antigens, Human Platelet/analysis
KW - Biosensing Techniques
KW - Blood Platelet Disorders/diagnosis
KW - Electrochemical Techniques
KW - Gold
KW - HLA Antigens/analysis
KW - Humans
KW - Isoantibodies/analysis
KW - Microelectrodes
KW - Protein Array Analysis/methods
KW - Sensitivity and Specificity
KW - Thrombocytopenia/diagnosis
U2 - 10.1016/j.bios.2012.04.021
DO - 10.1016/j.bios.2012.04.021
M3 - SCORING: Journal article
C2 - 22572157
VL - 36
SP - 207
EP - 211
JO - BIOSENS BIOELECTRON
JF - BIOSENS BIOELECTRON
SN - 0956-5663
IS - 1
ER -
