Kinetic Analysis of TNF Immune Complex by Using Surface Plasmon Resonance

Chem. J. Chinese Universities ›› 2002, Vol. 23 ›› Issue (3): 382.

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Kinetic Analysis of TNF Immune Complex by Using Surface Plasmon Resonance

YANG Fan¹, CUI Xiao-Qiang¹, YANG Xiu-Rong¹, CHEN Ting², LU Jun²

1. State Key Lab of Electroanalytical Chemistry, National Analytical Research Center of Electrochemistryand Spectroscopy, Changchun Institute of App lied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
2. Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China

Received:2000-12-20 Online:2002-03-24 Published:2002-03-24

Abstract

Abstract: The kinetic analysis of the interaction between tumor necrosis factor(TNF) and its monoclonal antibody was performed by surface plasmon resonance(SPR) technique. The monoclonal antibody was immobilized to the surface of CM5 sensor chip by amine coupling. TNFat different concentrations was injected across the mAb immobilized surface. The interaction was recorded in real time and could be seen on the sensorgram. One cycle, including association, dissociation and regeneration, lasted no more than 15min . The interaction results was evaluated using 1:1 Langmuir binding model. The kinetic rate constants were calculated to be: k_a=1.68×10³ L·mol^-1 ·s^-1 , k_d=1.73×10^-4 s^-1 , and the affinity constants K_A=9.7×10⁶ L·mol^-1 , K_D=1.03×10^-7 mol·L^-1 . The χ² was 3.47, which showed that the interaction is consistent with the 1:1 model. We can see from the results that although there are two binding sites in one mAb molecule, TNFreacts with each site in an independent and noncooperative manner.

Key words: Surface plasmon resonance, Tumor necrosis factor, Kinetic analysis

CLC Number:

O657

TrendMD:

YANG Fan, CUI Xiao-Qiang, YANG Xiu-Rong, CHEN Ting, LU Jun. Kinetic Analysis of TNF Immune Complex by Using Surface Plasmon Resonance[J]. Chem. J. Chinese Universities, 2002, 23(3): 382.

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Kinetic Analysis of TNF Immune Complex by Using Surface Plasmon Resonance

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