title: Evaluation and optimization of the metal-binding properties of a complex ligand for immobilized metal affinity chromatography.
abstract: The simultaneous determination of two binding parameters for metal ions on an immobilized metal affinity chromatography column was performed by frontal chromatography. In this study, the binding parameters of Cu(2+) to l-glutamic acid were measured, the metal ion-binding characteristics of the complex ligand were evaluated. The linear correlation coefficients were all greater than 99%, and the relative standard deviations of two binding parameters were 0.58 and 0.059%, respectively. The experiments proved that the frontal chromatography method was accurate, reproducible, and could be used to determine the metal-binding parameters of the affinity column. The effects of buffer pH, type, and concentration on binding parameters were explored by uniform design experiment. Regression, matching and residual analyses of the models were performed. Meanwhile, the optimum-binding conditions of Cu(2+) on the l-glutamic acid-silica column were obtained. Under these binding conditions, observations and regression values of two parameters were similar, and the observation values were the best. The results demonstrated that high intensity metal affinity column could be effectively prepared by measuring and evaluating binding parameters using frontal chromatography combined with a uniform design experiment. The present work provided a new mode for evaluating and preparing immobilized metal affinity column with good metal-binding behaviors.
authors.affiliation: Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, School of Chemical Engineering, Northwest University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Chen
authors.affiliation: Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, School of Chemical Engineering, Northwest University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Rong
authors.initials: R
authors.lastname: Li
authors.affiliation: Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, School of Chemical Engineering, Northwest University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Shiyu
authors.initials: S
authors.lastname: Li
authors.affiliation: Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, School of Chemical Engineering, Northwest University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.affiliation: School of Natural and Applied Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Kaidi
authors.initials: K
authors.lastname: Yang
authors.affiliation: Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, School of Chemical Engineering, Northwest University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Guoliang
authors.initials: G
authors.lastname: Chen
authors.affiliation: Chemical Engineering Research Center of the Ministry of Education for Advanced Use Technology of Shanbei Energy, School of Chemical Engineering, Northwest University, Xi'an, Shaanxi Province, P. R. China.
authors.forename: Xiaoxun
authors.initials: X
authors.lastname: Ma
keywords: Binding parameters; Frontal chromatography; Immobilized metal affinity chromatography; Uniform design