Douglas J. Cecchini

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.

A Sensitive Bioassay for a Standard of Recombinant Interleukin-1α

Abstract A sensitive bioassay for recombinant interleukin-1α (rIL-1α) has been developed. The assay is based on prostaglandin E2 (PGE2) production by fibroblasts following stimulation with rIL-1α. The stimulated PGE2 levels are determined by radioimmunoassay. Three human fibroblast cell lines, WI38, HEL299 and HS68, were evaluated. The HS68 cells gave the most sensitive and reproducible response. The results from six experiments established that 400 ± 300 femtograms of rIL-1α stimulated a two-fold increase in PGE2 concentration over the background level.