Preparation of magnetic immobilized metal affinity separation media and its use in the isolation of proteins

被引:85
作者
Abudiab, T [1 ]
Beitle, RR [1 ]
机构
[1] Univ Arkansas, Dept Chem Engn, Bell Engn Ctr 3202, Fayetteville, AR 72701 USA
关键词
immobilized metal affinity chromatography; magnetic media; stationary phases; LC; proteins;
D O I
10.1016/S0021-9673(97)00959-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A new method of pseudobiospecific protein isolation is developed and tested, which employs both metal affinity and magnetism as the basis for isolation. The chelating group iminodiacetic acid (IDA) has been coupled to the surface of magnetic agarose, and when charged with metal ions (Cu2+ or Zn2+) is capable of binding model proteins which display metal affinity, and of separating protein mixtures. Magnetic properties of the medium facilitated the batch recovery of the adsorbent, as losses are minimized by concentrating and retaining the separation medium with the aid of a magnet. Model proteins were used to characterize protein adsorption, capacity, and stability of IDA magnetic agarose. Recovery from a cell lysate was demonstrated by protein isolation from extracts of E. coli containing a target protein. Overall, this study effectively illustrates the engineering of separation media which combine several desired properties for the development of a new branch of metal affinity-based bioseparation. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:211 / 217
页数:7
相关论文
共 46 条
[1]   PURIFICATION OF COMMERCIAL HUMAN-ALBUMIN ON IMMOBILIZED IDA-NI2+ [J].
ANDERSSON, L ;
SULKOWSKI, E ;
PORATH, J .
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS, 1987, 421 (01) :141-146
[2]  
Andersson L, 1991, Bioseparation, V2, P15
[3]   METAL-AFFINITY SEPARATIONS - A NEW DIMENSION IN PROTEIN PROCESSING [J].
ARNOLD, FH .
BIO-TECHNOLOGY, 1991, 9 (02) :151-156
[4]  
BEITLE R, 1992, NEW DEV BIOSEPARATIO
[5]   ONE-STEP PURIFICATION OF A MODEL PERIPLASMIC PROTEIN FROM INCLUSION-BODIES BY ITS FUSION TO AN EFFECTIVE METAL-BINDING PEPTIDE [J].
BEITLE, RR ;
ATAAI, MM .
BIOTECHNOLOGY PROGRESS, 1993, 9 (01) :64-69
[6]   FRACTIONATION OF BASIC NUCLEAR PROTEINS OF HUMAN SPERM BY ZINC CHELATE AFFINITY-CHROMATOGRAPHY [J].
BIANCHI, F ;
ROUSSEAUXPREVOST, R ;
SAUTIERE, P ;
ROUSSEAUX, J .
JOURNAL OF CHROMATOGRAPHY, 1990, 518 (01) :123-134
[7]   IMMOBILIZED METAL-ION AFFINITY PARTITIONING, A METHOD COMBINING METAL PROTEIN-INTERACTION AND PARTITIONING OF PROTEINS IN AQUEOUS 2-PHASE SYSTEMS [J].
BIRKENMEIER, G ;
VIJAYALAKSHMI, MA ;
STIGBRAND, T ;
KOPPERSCHLAGER, G .
JOURNAL OF CHROMATOGRAPHY, 1991, 539 (02) :267-277
[8]   CONTINUOUS AFFINITY-CHROMATOGRAPHY USING A MAGNETICALLY STABILIZED FLUIDIZED-BED [J].
BURNS, MA ;
GRAVES, DJ .
BIOTECHNOLOGY PROGRESS, 1985, 1 (02) :95-103
[9]   STRUCTURAL STUDIES OF A LIQUID-FLUIDIZED MAGNETICALLY STABILIZED BED [J].
BURNS, MA ;
GRAVES, DJ .
CHEMICAL ENGINEERING COMMUNICATIONS, 1988, 67 :315-330
[10]   DRIED CALCIUM ALGINATE MAGNETITE SPHERES - A NEW SUPPORT FOR CHROMATOGRAPHIC SEPARATIONS AND ENZYME IMMOBILIZATION [J].
BURNS, MA ;
KVESITADZE, GI ;
GRAVES, DJ .
BIOTECHNOLOGY AND BIOENGINEERING, 1985, 27 (02) :137-145