Enhancement of immunoaffinity chromatography processes using polyclonal antibodies and high pressure elution

Enhanced elution of polyclonal-based immunoaffinity complexes by high pressure manipulations were examined. Although antigen (product) elution using polyclonal antibodies often proves difficult, it was demonstrated that high pressures (2 kbar) may dissociate a polyclonal antibody-antigen complex. Using this feature of high pressure, a unique high pressure cell was custom designed and built to accomplish the recovery of protein products from immunoadsorbents. The results from this work demonstrate that moderate recoveries using a polyclonal antibovine serum albumin (BSA) - BSA antigen system were achieved at 1724 bar using pressure elution. In contrast, low recoveries were achieved using a polyclonal anti-bovine immunoglobulin G (IgG) - bovine IgG system under the same pressure elution conditions. These findings indicate the need to screen antigen-antibody systems being considered for immunoaffinity chromatography operations employing pressure elution. Fourier transform infrared spectroscopy has been demonstrated to be a potentially useful approach to accomplish this.