Optimization of monoclonal antibody immobilization on hydrazide-preactivated hollow fiber membrane.

Immobilization of an IgG1 monoclonal antibody (MAb) was optimized using a unique hydrazide-preactivated hydrophilic hollow fiber membrane as the support matrix. Modules containing 0.42 milliliters of membrane volume (mlmv) were offered varying amounts of purified MAb. The highest immobilization efficiency on the hollow fiber membranes was 88% at a MAb loading concentration of 0.35 mg/ml. The optimum range of MAb concentrations to achieve the best immobilization efficiency was 0.18-0.45 mg/ml. A larger module containing 9.7 mlmv immobilized greater than 3.0 mg MAb/mlmv at an efficiency of greater than 90%. The total amount of MAb immobilized on the membranes within each module was in direct proportion to the total amount of membrane volume. Preliminary data suggest the optimized immunoaffinity hollow fiber membrane matrix produced in this study is stable and can achieve a product capacity of greater than 2.0 mg/mlmv. In concert with an automated fluid handling system, such as the TRIO(TM) Bioprocessing System, rapid accurate information can be easily generated on process parameters and scale-up considerations where an immunoaffinity step is included in the downstream purification protocol.