Affinity chromatography of trypsin using chitosan as ligand support

被引:39
作者
Shi, YC [1 ]
Jiang, YM [1 ]
Sui, DX [1 ]
Li, YL [1 ]
Chen, TA [1 ]
Ma, L [1 ]
Ding, ZT [1 ]
机构
[1] JILIN UNIV,NATL LAB ENZYME ENGN,CHANGCHUN 130023,PEOPLES R CHINA
关键词
affinity adsorbents; stationary phases; LC; trypsin; chitosan; ovomucoid; proteins;
D O I
10.1016/0021-9673(96)00260-9
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Chitosan beads were prepared for use as affinity adsorbent carrier. The affinity ligand, chicken ovomucoid, was immobilized on the chitosan via a cross-linker, glutaraldehyde. The results showed that 60 mg chicken ovomucoid could be immobilized on 1 g chitosan, and the maximum binding capacity for trypsin was about 8 . 10(4) U/g dry adsorbent. The procedure for preparing the chitosan-based affinity adsorbents was much safer and simpler than when a Sepharose-based matrix was the support. Columns packed with the affinity adsorbents were employed for trypsin chromatography. The experimental results revealed that the affinity adsorbents possessed good mechanical strength and storage stability and could be also operated repeatedly. Chitosan was suitable for use as an affinity adsorbent support for laboratory-scale and large-scale purification.
引用
收藏
页码:107 / 112
页数:6
相关论文
共 12 条
[1]  
Broussignac P., 1968, CHIM IND GEN CHIM, V99, P1241
[2]   PROTEASE IMMOBILIZATION ONTO POROUS CHITOSAN BEADS [J].
HAYASHI, T ;
IKADA, Y .
JOURNAL OF APPLIED POLYMER SCIENCE, 1991, 42 (01) :85-92
[3]   SYNTHESIS AND EVALUATION OF CHITOSAN-BASED, AFFINITY-CHROMATOGRAPHY MATERIALS [J].
HOLME, KR ;
HALL, LD ;
ARMSTRONG, CR ;
WITHERS, SG .
CARBOHYDRATE RESEARCH, 1988, 173 (02) :285-291
[4]  
KASSELL B, 1970, METHOD ENZYMOL, V19, P890
[5]  
LINEWEAVER H, 1947, J BIOL CHEM, V171, P565
[6]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[7]   PREPARATIVE AFFINITY-CHROMATOGRAPHY OF PROTEINS [J].
NARAYANAN, SR .
JOURNAL OF CHROMATOGRAPHY A, 1994, 658 (02) :237-258
[8]  
RICK W, 1965, METHOD ENZYMAT AN, P800
[9]   DETERMINATION OF THE VISCOMETRIC CONSTANTS FOR CHITOSAN [J].
ROBERTS, GAF ;
DOMSZY, JG .
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 1982, 4 (06) :374-377
[10]   AFFINITY-PRECIPITATION USING CHITOSAN AS LIGAND CARRIER [J].
SENSTAD, C ;
MATTIASSON, B .
BIOTECHNOLOGY AND BIOENGINEERING, 1989, 33 (02) :216-220