Some parameters of affinity chromatography in the purification of antibody against Naja naja siamensis toxin 3.
Some parameters in the purification of antibodies against Naja naja siamensis toxin 3 by affinity chromatography were studied on toxin Sepharose, toxin-succinylaminoethyl Sepharose, toxin-albumin Sepharose and toxin succinylaminoethyl Biogel adsorbents. Immunologically pure antibody with 10-12-fold increase in potency was obtained by chromatography of horse refined globulin on all these adsorbents. The maximum antibody binding capacities were higher for adsorbents containing linear spacers but represented only 7-16% of the theoretical binding capacity. The operational half-lives of the adsorbents ranged from 19 to 108 days with toxin-albumin Sepharose showing the highest stability. The recovery of neutralizing capacity was about 30-356% for any of the 4 adsorbents. It is concluded that improvements regarding the antibody binding capacity and the recovery of neutralizing capacity should be made before large scale purification of antibody for therapeutic purposes can be attempted.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000997:Antivenins / Q000302:isolation & purification*; D001666:Binding Sites, Antibody; D002846:Chromatography, Affinity; D004546:Elapid Venoms / Q000276:immunology*; D006207:Half-Life; D006736:Horses; D007164:Immunosorbents / Q000494:pharmacology; D009500:Neutralization Tests; D012685:Sepharose / Q000494:pharmacology; D012712:Serum Globulins / Q000276:immunology
D000906:Antibodies; D000997:Antivenins; D004546:Elapid Venoms; D007164:Immunosorbents; D012712:Serum Globulins; D012685:Sepharose