Polystyrene as an affinity chromatography matrix for the purification of antibodies.
Affinity chromatography is used for the purification of diagnostic polyclonal antibodies in order to ensure specificity. Most commonly, activated bead-formed agarose or its derivatives are used as gel matrices. Alternative matrix materials have been described, but as yet they do not appear to offer important advantages. In this study, pulverized polystyrene (PS 158K, BASF, Mannheim, Germany) was used as a solid phase for the immobilisation of bovine immunoglobulins (Ig). Affinity chromatography was performed using these coated polystyrene beads as the column matrix material in the purification of anti-bovine Ig. The polystyrene binding capacity for the different bovine Ig classes was compared using the Mancini single radial immunodiffusion technique, and ELISA procedures were used to monitor the antibody reactivity of purified and unpurified antibodies. The degree of purification was comparable to the most commonly used procedure using gel matrices from activated bead-formed agarose (e.g. CNBr-activated Sepharose 4B, Pharmacia/LKB Biotechnology, Uppsala, Sweden), but the antibody yield per ml column volume was distinctly lower. In order to raise the yield, such polystyrene bead columns with immobilized antigen can be re-used without loss of activity or larger column volumes can be used to raise the binding capacity. The polystyrene material is quite durable, chemically and immunologically inert and has a long shelf life. We conclude that polystyrene based affinity chromatography is efficient, simple and cheap.
Staak
C
C
BGVV, Berlin, Germany.
Salchow
F
F
Clausen
P H
PH
Luge
E
E
D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000302:isolation & purification*; D000918:Antibody Specificity; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D007136:Immunoglobulins / Q000737:chemistry; D007164:Immunosorbents; D011137:Polystyrenes; D011485:Protein Binding
D000906:Antibodies; D007136:Immunoglobulins; D007164:Immunosorbents; D011137:Polystyrenes