Kinetic analysis of the interaction between amphotericin B and human serum albumin using surface plasmon resonance and fluorescence spectroscopy.
The binding interaction between amphotericin B and human serum albumin (HSA) has been studied using surface plasmon resonance (SPR) spectroscopy combined with a fluorescence quenching method to confirm the binding kinetic results. In this paper, the SPR method used to study the drug-protein interaction has been described in detail. The association rate constant, dissociation rate constant and the equilibrium association constant of amphotericin B binding to HSA were obtained using this method. To confirm the feasibility of the SPR method, a fluorescence quenching method was performed to obtain the equilibrium constant. In order to obtain more accurate results, experiment design was used to optimize the fluorescence quenching process. The two equilibrium association constants obtained using the two methods were 4.017 x 10(4) M(-1) (SPR) and 3.656 x 10(4) M(-1) (fluorescence quenching method) respectively.
Zhou
Bo
B
State Key Laboratory of Virology, College of Chemistry and Molecular Science, Wuhan University, Wuhan 430072, P R China.
Li
Ran
R
Zhang
Yue
Y
Liu
Yi
Y
D000666:Amphotericin B / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D012709:Serum Albumin / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013455:Sulfur / Q000737:chemistry; D020349:Surface Plasmon Resonance
D012709:Serum Albumin; D013455:Sulfur; D006046:Gold; D000666:Amphotericin B