Quartz crystal biosensor for real-time kinetic analysis of interaction between human TNF-α and monoclonal antibodies

• Published in: Sensors and actuators. B, Chemical Vol. 99; no. 2; pp. 416 - 424
• Main Authors: Liu, Yang, Zhang, Wei, Yu, Xiao, Zhang, Hongwu, Zhao, Rui, Shangguan, DiHua, Li, Yan, Shen, Beifen, Liu, Guoquan
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.05.2004
• Subjects: Algorithms; Biosensors; Electrodes; ELISA; Enzyme-linked immunosorbent assay; Genetic algorithm; Human TNF-α; Kinetic analysis; Monoclonal antibodies; Q1; Quartz; Quartz crystal biosensor; Tumor necrosis factor- alpha; Human TNF-α; Kinetic analysis; Genetic algorithm; Quartz crystal biosensor; ELISA
• ISSN: 0925-4005; 1873-3077
• DOI: 10.1016/j.snb.2003.12.010

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.