Quartz crystal biosensor for real-time kinetic analysis of interaction between human TNF-α and monoclonal antibodies
Quartz crystal microbalance (QCM) biosensor integrated in a flow injection analysis (FIA) system was used for the real-time investigation of the interaction between human TNF-α (hTNF-α) and its monoclonal antibodies (mAbs). Biologically active hTNF-α trimer was immobilized on the gold electrodes of...
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Published in | Sensors and actuators. B, Chemical Vol. 99; no. 2; pp. 416 - 424 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Elsevier B.V
01.05.2004
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Subjects | |
Online Access | Get full text |
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Summary: | Quartz crystal microbalance (QCM) biosensor integrated in a flow injection analysis (FIA) system was used for the real-time investigation of the interaction between human TNF-α (hTNF-α) and its monoclonal antibodies (mAbs). Biologically active hTNF-α trimer was immobilized on the gold electrodes of the piezoelectric crystals and the binding interactions of the two immobilized antigen with two anti-hTNF-α monoclonal antibodies (Z8 and Z12) were followed as changes in the resonant frequency of the modified crystals. The resultant sensorgrams were rapidly analyzed by using an in-house kinetic analysis software based on genetic algorithm (GA) to derive both the kinetic rate constants (
k
ass and
k
diss) and equilibrium dissociation constant (
K
D) for antigen–antibody interactions. For the interactions,
K
D were 2.31×10
−7 and 5.42×10
−7
M for Z8 and Z12, respectively. The generally good agreement found between biosensor data and the affinities known from conventional equilibrium enzyme-linked immunosorbent assay (ELISA) validates this analytical method. The study shows the potential usefulness of QCM biosensor technology for biomolecular interaction analysis. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 |
ISSN: | 0925-4005 1873-3077 |
DOI: | 10.1016/j.snb.2003.12.010 |