Optimised affinity purification of polyclonal antibodies from hyper immunised ovine serum using a synthetic Protein A adsorbent, MAbsorbent A2P.
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 A2P was shown to bind approximately 27 mg mL(-1) of ovine immunoglobulin from undiluted serum, with eluted IgG purities of >95%, minor levels of albumin (approximately 1%) 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 A2P is a feasible alternative to Protein A chromatography or sodium sulphate precipitation for the initial capture of antibodies from undiluted serum.
Newcombe
Anthony R
AR
Process Development Group, Protherics Ltd., Blaenwaun, Ffostrasol, Llandysul, Wales SA44 5JT, UK. tony.newcombe@protherics.com
Cresswell
Chrissie
C
Davies
Susannah
S
Watson
Keith
K
Harris
Guy
G
O'Donovan
Kieran
K
Francis
Richard
R
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera / Q000276:immunology*; D007525:Isoelectric Focusing; D012756:Sheep; D013205:Staphylococcal Protein A / Q000737:chemistry*
D000906:Antibodies; D007106:Immune Sera; D013205:Staphylococcal Protein A