Simple affinity purification of antibodies using in vivo biotinylation of a fusion protein

被引:8
作者
Lesley, SA
Groskreutz, DJ
机构
[1] Promega Corporation, Madison, WI 53711
关键词
antibody purification; affinity purification; biotinylation; avidin; IgY;
D O I
10.1016/S0022-1759(97)00113-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We describe a method for affinity purification of antibodies using gene fusions to the antigen. Fusions are made to a protein domain which is recognized in vivo by biotin ligase and biotinylated at a unique position in the fusion protein. When expressed in E. coli, the fusion protein can be captured from a whole cell lysate using an avidin resin to generate an affinity column which is useful for antibody purification. The procedure is simple, rapid and does not require chemical biotinylation or prior purification of the target antigen. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:147 / 155
页数:9
相关论文
共 19 条
[1]   Structure of the biotinyl domain of acetyl-coenzyme A carboxylase determined by MAD phasing [J].
Athappilly, FK ;
Hendrickson, WA .
STRUCTURE, 1995, 3 (12) :1407-1419
[2]  
BERLINER E, 1994, J BIOL CHEM, V269, P8610
[3]   EXPRESSION, BIOTINYLATION AND PURIFICATION OF A BIOTIN-DOMAIN PEPTIDE FROM THE BIOTIN CARBOXY CARRIER PROTEIN OF ESCHERICHIA-COLI ACETYL-COA CARBOXYLASE [J].
CHAPMANSMITH, A ;
TURNER, DL ;
CRONAN, JE ;
MORRIS, TW ;
WALLACE, JC .
BIOCHEMICAL JOURNAL, 1994, 302 :881-887
[4]   PROPERTIES AND PURIFICATION OF AN ACTIVE BIOTINYLATED LACTOSE PERMEASE FROM ESCHERICHIA-COLI [J].
CONSLER, TG ;
PERSSON, BL ;
JUNG, H ;
ZEN, KH ;
JUNG, K ;
PRIVE, GG ;
VERNER, GE ;
KABACK, HR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (15) :6934-6938
[5]  
CRONAN JE, 1990, J BIOL CHEM, V265, P10327
[6]   INCREASE IN STABILITY OF AVIDIN PRODUCED BY BINDING OF BIOTIN - DIFFERENTIAL SCANNING CALORIMETRIC STUDY OF DENATURATION BY HEAT [J].
DONOVAN, JW ;
ROSS, KD .
BIOCHEMISTRY, 1973, 12 (03) :512-517
[7]  
FALL RR, 1973, J BIOL CHEM, V248, P2078
[8]  
Green N M, 1975, Adv Protein Chem, V29, P85, DOI 10.1016/S0065-3233(08)60411-8
[9]   AVIDIN .3. NATURE OF BIOTIN-BINDING SITE [J].
GREEN, NM .
BIOCHEMICAL JOURNAL, 1963, 89 (03) :599-&
[10]   THE MECHANISM OF BIOTIN-DEPENDENT ENZYMES [J].
KNOWLES, JR .
ANNUAL REVIEW OF BIOCHEMISTRY, 1989, 58 :195-221