Superparamagnetic adsorbents for high-gradient magnetic fishing of lectins out of legume extracts

被引:51
作者
Heeboll-Nielsen, A [1 ]
Dalkiær, M [1 ]
Hubbuch, JJ [1 ]
Thomas, ORT [1 ]
机构
[1] Tech Univ Denmark, Ctr Proc Biotechnol, Bioctr, DK-2800 Lyngby, Denmark
关键词
affinity separation; batch adsorption; concanavalin A; Lens culinaris agglutinin; high-gradient magnetic separation; hydrophobic charge induction; mixed-mode ligands;
D O I
10.1002/bit.20116
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
This work presents the development, testing, and application in high-gradient magnetic fishing of superparamagnetic supports for adsorption of lectins. Various approaches were examined to produce affinity, mixed mode, and hydrophobic charge induction type adsorbents. In clean monocomponent systems affinity supports created by direct attachment of glucose or maltose to amine-terminated iron oxide particles could bind concanavalin A at levels of up to approximate to 280 mg g(-1) support with high affinity (approximate to 1 muM dissociation constants). However, the best performance was delivered by adsorbents featuring coupled tentacular dextran chains displaying a maximum binding capacity of 238 mg g(-1) and a dissociation constant of 0.13 muM. Adsorbents derivatized with mixed mode or hydrophobic charge induction ligands likewise demonstrated very high capacities for both concanavalin A and Lens culinaris agglutinin (greater than or equal to 250 mg g(-1)) with dissociation constants in the micromolar range, though neither of these systems showed any selectivity for lectins in leguminous extracts. When the affinity supports were applied to carbohydrate containing legume extracts only the dextran-linked adsorbents supplied sufficient competition to dissolved sugars to selectively bind concanavalin A in an extract of jack beans. The dextran-linked supports were employed in a high-gradient magnetic fishing experiment, in which concanavalin A was purified to near homogeneity from a crude, unclarified extract of jack beans. (C) 2004 Wiley Periodicals, Inc.
引用
收藏
页码:311 / 323
页数:13
相关论文
共 32 条
[1]   Expanded-bed chromatography in primary protein purification [J].
Anspach, FB ;
Curbelo, D ;
Hartmann, R ;
Garke, G ;
Deckwer, WD .
JOURNAL OF CHROMATOGRAPHY A, 1999, 865 (1-2) :129-144
[2]   IMMOBILIZATION OF REDUCING OLIGOSACCHARIDES TO MATRICES BY A GLYCOSYLAMIDE LINKAGE [J].
BLOMBERG, L ;
WIESLANDER, J ;
NORBERG, T .
JOURNAL OF CARBOHYDRATE CHEMISTRY, 1993, 12 (03) :265-276
[3]   Hydrophobic charge induction chromatography: salt independent protein adsorption and facile elution With aqueous buffers [J].
Burton, SC ;
Harding, DRK .
JOURNAL OF CHROMATOGRAPHY A, 1998, 814 (1-2) :71-81
[4]   POLYPEPTIDE LIGATION OCCURS DURING POST-TRANSLATIONAL MODIFICATION OF CONCANAVALIN-A [J].
CARRINGTON, DM ;
AUFFRET, A ;
HANKE, DE .
NATURE, 1985, 313 (5997) :64-67
[5]  
EDELMAN GM, 1978, J BIOL CHEM, V253, P3016
[6]   TECHNIQUES AND INSTRUMENTATION FOR PREPARATIVE IMMUNOSORBENT SEPARATIONS [J].
EVELEIGH, JW .
JOURNAL OF CHROMATOGRAPHY, 1978, 159 (01) :129-145
[7]   Immobilization of pectin fragments on solid supports: Novel coupling by thiazolidine formation [J].
Guillaumie, F ;
Thomas, ORT ;
Jensen, KJ .
BIOCONJUGATE CHEMISTRY, 2002, 13 (02) :285-294
[8]   MAGNETIC SUPPORTS FOR IMMOBILIZED ENZYMES AND BIOAFFINITY ADSORBENTS [J].
HALLING, PJ ;
DUNNILL, P .
ENZYME AND MICROBIAL TECHNOLOGY, 1980, 2 (01) :2-10
[9]   Efficient inclusion body processing using chemical extraction and high gradient magnetic fishing [J].
Heeboll-Nielsen, A ;
Choe, WS ;
Middelberg, APJ ;
Thomas, ORT .
BIOTECHNOLOGY PROGRESS, 2003, 19 (03) :887-898
[10]  
HEEBOLLNIELSEN A, 2002, THESIS TU DENMARK LY