Optimised affinity purification of polyclonal antibodies from hyper immunised ovine serum using a synthetic Protein A adsorbent, MAbsorbento® A2P

被引:68
作者
Newcombe, AR [1 ]
Cresswell, C
Davies, S
Watson, K
Harris, G
O'Donovan, K
Francis, R
机构
[1] Prother Ltd, Proc Dev Grp, Llandysul SA44 5JT, Wales
[2] ProMet BioSci Ltd, Tech Support Grp, Cambridge CB4 07A, England
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2005年 / 814卷 / 02期
关键词
mimetic; protein A; chromatography; affinity purification; antibody; serum;
D O I
10.1016/j.jchromb.2004.10.027
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This report describes the applicability of a synthetic chromatography adsorbent for large-scale purification of polyclonal immunoglobulin G from hyper immunised ovine serum. Under optimised conditions, MAbsorbent((R)) A2P was shown to bind similar to27 mg mL(-1) of ovine immunoglobulin from undiluted serum, with eluted IgG purities of >95%, minor levels of albumin (similar to1%) and undetectable levels of leached ligand in the purified preparations. The results presented here indicate that the optimised affinity capture of immunoglobulin from ovine serum using MAbsorbent((R)) A2P is a feasible alternative to Protein A chromatography or sodium sulphate precipitation for the initial capture of antibodies from undiluted serum. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:209 / 215
页数:7
相关论文
共 20 条
[1]  
[Anonymous], 2002, RUL GUID PHARM MAN D
[2]  
BAKER RM, 1994, SEPARATION BIOTECHNO, V3, P427
[3]  
*CROFAB, 2001, MED LETT, V1107, P55
[4]   Fatty acid binding to human serum albumin: new insights from crystallographic studies [J].
Curry, S ;
Brick, P ;
Franks, NP .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 1999, 1441 (2-3) :131-140
[5]   PURIFICATION OF F(AB')2 ANTI-SNAKE VENOM BY CAPRYLIC-ACID - A FAST METHOD FOR OBTAINING IGG FRAGMENTS WITH HIGH NEUTRALIZATION ACTIVITY, PURITY AND YIELD [J].
DOSSANTOS, MC ;
LIMA, MRD ;
FURTADO, GC ;
COLLETTO, GMDD ;
KIPNIS, TL ;
DASILVA, WD .
TOXICON, 1989, 27 (03) :297-303
[6]   Toward a new cash cow [J].
Echelard, Y ;
Meade, H .
NATURE BIOTECHNOLOGY, 2002, 20 (09) :881-882
[7]  
FDA, 2004, COD FED REG CFR TITL
[8]   Factorial screening of antibody purification processes using three chromatography steps without protein A [J].
Follman, DK ;
Farner, RL .
JOURNAL OF CHROMATOGRAPHY A, 2004, 1024 (1-2) :79-85
[9]   New method for the selective capture of antibodies under physiolgical conditions [J].
Guerrier, L ;
Girot, P ;
Schwartz, W ;
Boschetti, E .
BIOSEPARATION, 2000, 9 (04) :211-221
[10]   Antibody manufacture in transgenic animals and comparisons with other systems [J].
Houdebine, LM .
CURRENT OPINION IN BIOTECHNOLOGY, 2002, 13 (06) :625-629