Method for removal of antibody cross-reacting albumin from polyclonal antibody

摘要

PURPOSE: An eliminating process for cross-reaction antibody with albumin from polyclonal antibody is provided, which can be used to enhance specificity of polyclonal antibody to blood protein that does not adsorbed to Cibacron Blue 3FG including 2-macroglobulin and heptoglobin. CONSTITUTION: The eliminating process for cross-reaction antibody with albumin from polyclonal antibody comprises the steps of: preparing 2-macroglobulin, and processing 50% ammonium sulfate and anion exchange resin or protein-A cepharose column chromatography to get polyclonal antibody to 2-macroglobulin; equilibrating Cibacron Blue 3FG cepharose gel(Bio-Rad) with phosphate buffer solution, passing this with raw mouse blood, and washing un-adsorbed protein completely with sodium chloride and phosphate buffer solution(in this step, albumin is adsorbed); passing the refined polyclonal antibody to 2-macroglobulin through the Cibacron Blue 3FG cepharose gel column(cross-reaction antibody with albumin remains on column and other antibody is dissolved out); and concentrating the dissolved antibody.