Separation of MBP fusion proteins through affinity membranes

被引:12
作者
Cattoli, F [1 ]
Sarti, GC [1 ]
机构
[1] Univ Bologna, Dipartimento Ingn Chim Mineraria & Tecnol Ambient, I-40136 Bologna, Italy
关键词
D O I
10.1021/bp010119r
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Cellulose microporous membranes have been modified in order to obtain a stationary phase specific for the recovery of a class of fusion proteins containing the maltose binding protein domain, through affinity chromatography separations. The feasibility of a single step separation process for the recovery of large amounts of the desired product has been considered. To that purpose, a preparative scale module has been realized, suitable for flat sheet membranes. The affinity matrix used proved to be highly selective toward the fusion proteins examined. The binding capacity determined is comparable with the nominal binding capacity of commercially available supports. The influence of the relevant working parameters, such as flow rate, on the performances of the recovery process has been studied.
引用
收藏
页码:94 / 100
页数:7
相关论文
共 24 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   FAST PROTEIN CHROMATOGRAPHY ON ANALYTICAL AND PREPARATIVE SCALE USING MODIFIED MICROPOROUS MEMBRANES [J].
BRIEFS, KG ;
KULA, MR .
CHEMICAL ENGINEERING SCIENCE, 1992, 47 (01) :141-149
[3]  
CALCATERRA SL, 2001, THESIS U BOLOGNA
[4]  
CATTOLI F, 2000, THESIS U BOLOGNA
[5]  
CATTOLI F, IN PRESS SEP SCI TEC
[6]  
CATTOLI F, 2001, Patent No. 2001A00058
[7]   Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element [J].
Chong, SR ;
Mersha, FB ;
Comb, DG ;
Scott, ME ;
Landry, D ;
Vence, LM ;
Perler, FB ;
Benner, J ;
Kucera, RB ;
Hirvonen, CA ;
Pelletier, JJ ;
Paulus, H ;
Xu, MQ .
GENE, 1997, 192 (02) :271-281
[8]  
GUAN C, 1988, GENE, V67, P21
[9]   ACTIVE-TRANSPORT OF MALTOSE IN ESCHERICHIA-COLI-K12 - INVOLVEMENT OF A PERIPLASMIC MALTOSE BINDING-PROTEIN [J].
KELLERMANN, O ;
SZMELCMAN, S .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1974, 47 (01) :139-149
[10]  
KELLERMANN OK, 1982, METHOD ENZYMOL, V90, P459