Affinity chromatography of trypsin using chitosan as ligand support.
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 l 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.
Shi
Y C
YC
Laboratory of Enzyme Engineering, Agricultural College, Yangzhou University, China.
Jiang
Y M
YM
Sui
D X
DX
Li
Y L
YL
Chen
T
T
Ma
L
L
Ding
Z T
ZT
D000327:Adsorption; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000378:metabolism; D001704:Biopolymers / Q000737:chemistry*; D002645:Chickens; D002686:Chitin / Q000031:analogs & derivatives* / Q000737:chemistry; D048271:Chitosan; D002846:Chromatography, Affinity / Q000379:methods*; D002848:Chromatography, DEAE-Cellulose; D002853:Chromatography, Liquid / Q000295:instrumentation; D005976:Glutaral / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008024:Ligands; D008863:Microspheres; D010059:Ovomucin / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D014357:Trypsin / Q000302:isolation & purification* / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000737:chemistry*
D001704:Biopolymers; D008024:Ligands; D014361:Trypsin Inhibitors; D002686:Chitin; D010059:Ovomucin; D048271:Chitosan; D001120:Arginine; C009727:benzoylarginine ethyl ester; D014357:Trypsin; D005976:Glutaral