Utility of Polycation-Treated Filters for the Assay of Receptors for VIP

Abstract

This study examined the utility of four polycationic agents for treating glass fibre filters used in the receptor binding assay for vasoactive intestinal peptide (VIP). Polyethylenimine (PEI), polybrene, protamine and methylated bovine serum albumin proved satisfactory in terms of low filter binding of free radioligand and retention of membrane-bound radioligand. Their performance was superior or comparable to untreated Millipore EGWP cellulose acetate filters which we had previously utilized but which are no longer manufactured. The results with polycations indicate the importance of ionic interactions between filter, biological membranes and radioligand in determining the performance of a filtration assay for radioligand-receptor binding. At a practical level, PEI has the disadvantage of potential toxicity. The satisfactory performance of the other polycations indicates that they provide safer alternatives to PEI for filtration assay of the VIP receptor and possibly receptors for other basic ligands.