Selection of ligands for affinity chromatography using quartz crystal   biosensor

doi:10.1021/ac050026e

CORC > 北京大学 > 化学与分子工程学院

	Selection of ligands for affinity chromatography using quartz crystal biosensor
	Liu, Y ; Tang, XL ; Liu, F ; Li, K
刊名	analytical chemistry
	2005
关键词	SALT-INDUCED IMMOBILIZATION SURFACE-PLASMON RESONANCE TIME KINETIC-ANALYSIS HUMAN SERUM-ALBUMIN COMBINATORIAL LIBRARIES MOLECULAR RECOGNITION LYSOZYME SEPARATION MASS-SPECTROMETRY IMMUNOGLOBULIN-G PURIFICATION
DOI	10.1021/ac050026e
英文摘要	This paper described a new strategy for rapid selecting ligands for application in affinity chromatography using a quartz crystal microbalance (QCM) biosensor. An aminoglycoside antibiotic drug, kanamycin, (KM), was immobilized on the gold electrodes of the QCM sensor chip. The binding interactions of the immobilized KM with various proteins in solution were monitored as the variations of the resonant frequency of the modified sensor. Such a rapid screen analysis of interactions indicated clearly that KM-immobilized sensor showed strong specific interaction only with lysozyme (LZM). The resultant sensorgrams were rapidly analyzed by using a kinetic analysis software based on a genetic algorithm to derive both the kinetic rate constants (k(ass) and k(diss)) and equilibrium dissociation constants (K-D) for LZM-KM interactions. The immobilized KM showed higher affinity to LZM with a dissociation constant on the order of 10(-5) M, which is within the range of 10(-4)-10(-8) M and suitable for an affinity ligand. Therefore, KM was demonstrated for the first time as a novel affinity ligand for purification of LZM and immobilized onto the epoxy-activated silica in the presence of a high potassium phosphate concentration. The KM immobilized affinity column has proved useful for a very convenient purification of LZM from chicken egg white. The purity of LZM obtained was higher than 90%, as determined by densitometric scanninng of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified fraction. These results confirmed that the selected KM ligand is indeed a valuable affinity ligand for purification of LZM. The new screening strategy based on a QCM biosensor is expected to be a promising way for rapid selecting specific ligands for purifying other valuable proteins.; Chemistry, Analytical; SCI(E); EI; PubMed; 25; ARTICLE; 13; 4248-4256; 77
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/199301]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Liu, Y,Tang, XL,Liu, F,et al. Selection of ligands for affinity chromatography using quartz crystal biosensor[J]. analytical chemistry,2005.
APA	Liu, Y,Tang, XL,Liu, F,&Li, K.(2005).Selection of ligands for affinity chromatography using quartz crystal biosensor.analytical chemistry.
MLA	Liu, Y,et al."Selection of ligands for affinity chromatography using quartz crystal biosensor".analytical chemistry (2005).