Purification of fibrin-specific monoclonal antibody from ascites fluid by preparative isoelectric focusing.
With the increased use of site-directed monoclonal antibodies (MAbs) for the detection and localization of antigens (Ags) in vivo, an efficient, noninjurious MAb purification procedure is essential [1-9]. In this study MAb was purified by both immunoaffinity chromatography (IAFC) and preparative isoelectric focusing (PIEF). For anti-fibrin MAb purified by each method, the purification efficiency, blood clearance, and in vivo localization in thrombi were compared. Blood clearance rates and in vivo localization were determined by a double isotope assay: affinity-purified MAb was labeled with I-125, and PIEF-purified MAb was labeled with I-131. Both were injected intravenously into a dog with an experimentally induced femoral vein thrombus. MAb purified by PIEF had a slightly longer biological T 1/2 resulting in comparably higher thrombus localization. Due to its efficiency, PIEF warrants consideration when purified MAb is desired.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001202:Ascitic Fluid / Q000276:immunology; D002846:Chromatography, Affinity; D005337:Fibrin / Q000276:immunology*; D005779:Immunodiffusion; D007525:Isoelectric Focusing / Q000379:methods*; D051379:Mice
D000911:Antibodies, Monoclonal; D005337:Fibrin