Tresyl activation of a hydroxyalkyl ligand for coupling to a hydrazide gel: stable immobilization of T-2 toxin for affinity purification of T-2 antibody.
A stable T-2 hydrazide gel is prepared by activating T-2 toxin with tresyl chloride followed by coupling to agarose-adipic acid hydrazide. Utilized as an affinity chromatography column, this T-2 hydrazide gel purifies a monoclonal antibody for T-2 in high yield directly from ascites fluid. Specific antibody trapped on the column is eluted either with excess T-2 or at pH 11.6. Much less successful are two other T-2 affinity columns that were prepared and evaluated: T-2 bovine serum albumin Affi-Gel 15 and T-2 hexylamine Sepharose.
D000906:Antibodies / Q000302:isolation & purification*; D001711:Biotransformation / Q000187:drug effects; D002846:Chromatography, Affinity; D005782:Gels; D006834:Hydrazines; D008024:Ligands; D011485:Protein Binding / Q000187:drug effects; D012685:Sepharose; D012717:Sesquiterpenes / Q000276:immunology*; D013450:Sulfones / Q000494:pharmacology*; D013605:T-2 Toxin / Q000276:immunology*
D000906:Antibodies; D005782:Gels; D006834:Hydrazines; D008024:Ligands; D012717:Sesquiterpenes; D013450:Sulfones; C032605:2,2,2-trifluoroethanesulfonyl chloride; D012685:Sepharose; D013605:T-2 Toxin