A glass-fiber filtration assay for solution phase hapten-antibody binding. Effect of surface treatment with a polycation.
Radioligands bound to murine monoclonal and polyclonal antibodies in solution are rapidly and effectively separated from free ligand by filtration through either untreated or polyethylenimine (PEI)-pretreated glass-fiber filters. Ligand binding to selected anti-morphine monoclonal antibodies, determined by filtration through untreated filters, was significantly greater (1.5-3.0-fold) than the binding activities obtained by gel filtration. However, radioactivity retained by filters that were pretreated with PEI (pH 4) was essentially the same as that obtained using the Sephadex G-25 short-column assay. The pH dependence of the retention on the coated fibers suggests that the mechanism of binding of antibodies to glass is different from that to the PEI-treated surfaces. The quantitative aspects of the assay are reported. The method should prove useful where quantitative, rapid and inexpensive binding radioassays need to be performed.
Miller
A
A
Department of Biochemistry, University of Connecticut Health Center, Farmington 06032.
Glasel
J A
JA
D000911:Antibodies, Monoclonal / Q000378:metabolism; D000936:Antigen-Antibody Complex; D005139:Eyeglasses; D005374:Filtration; D006241:Haptens / Q000302:isolation & purification* / Q000378:metabolism; D007097:Imines; D008024:Ligands; D009020:Morphine / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D011073:Polyamines; D000071228:Polyelectrolytes; D011095:Polyethylenes; D011108:Polymers; D012995:Solubility
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006241:Haptens; D007097:Imines; D008024:Ligands; D009020:Morphine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011073:Polyamines; D000071228:Polyelectrolytes; D011095:Polyethylenes; D011108:Polymers