Development and monitoring of purification process for nerve growth factor fusion antibody

Abstract A purification process and an on-line process monitoring procedure for a human nerve growth factor (NGF) fusion antibody (NAK) was developed. The NAK protein was produced with a recombinant technique by an adherent Chinese hamster ovary cell line in a tissue culture system supplemented with 5–10% fetal bovine serum, then purified from tissue culture supernatant. The overall purity goal for the process was 90%. A capture step was developed and optimized on POROS 50A which served as a purification and concentration step for the NAK product. A second purification step to remove contaminating bovine IgGs was developed and optimized first on a POROS HS/M cation-exchange column using 20-μm particles. The process was then scaled to a POROS 50 HS (50-μm particle) column. Once an optimized process was developed the capture step on POROS 50A was scaled from an 8-ml column to a 0.3-1 column on PerSeptive Biosystems AutoPilot system.