Purification of antibodies using the synthetic affinity ligand absorbent MAbsorbent A2P.
A protocol for the purification of polyclonal antibodies from ovine serum using the synthetic protein A absorbent MAbsorbent A2P is described. Clarified serum is loaded directly onto the affinity column without prior adjustment and albumin and unwanted serum components are washed from the column using a sodium octanoate buffer before elution of bound antibodies. MAbsorbent A2P was shown to bind approximately 27 mg ml(-1) of polyclonal immunoglobulin under overloading conditions, with eluted IgG purities of >90% and minor levels of albumin (approximately 1%). The anticipated time required to complete the purification protocol is 6-7 h. Although the protocol is similar to methods utilized for antibody purification using chromatography with protein A derived from the cell wall of the microorganism Staphylococcus aureus or protein G from Streptococcus as the affinity ligands, affinity absorbents based on synthetic ligands offer a number of advantages to compounds derived from biological sources, in particular robustness, relatively low cost, ease of sanitization and, in principle, lack of biological contamination.
Chhatre
Sunil
S
Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK.
Titchener-Hooker
Nigel J
NJ
Newcombe
Anthony R
AR
Keshavarz-Moore
Eli
E
D000042:Absorption; D000818:Animals; D000906:Antibodies / Q000097:blood* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D004586:Electrophoresis; D005374:Filtration; D008024:Ligands; D012756:Sheep
D000906:Antibodies; D000911:Antibodies, Monoclonal; D008024:Ligands