Process Development for the Production of a Monoclonal Antibody Against Francisella Tularensis.

摘要

Antibodies are essential components in biosensors and enable the detection of biological warfare agents with both sensitivity and selectivity. A biomanufacturing process was developed for the production of a monoclonal antibody (mAb) specific for detecting the Frencisella tularensls organism in the environment. This report describes a process for the production of 12 g of monocional antibody from an in vitro call culture system. Hybridoma calls were grown in 2-L gas permeable call culture bags. The numbers of both total and live calls in culture media were measured throughout the process and correlated with antibody concentration over time. Antibody concentrations in samples collected during culture were monitored and during the purification process were analyzed by analytical high-performance liquid chromatography. Antibodies were purified from spent culture media, using a single-step Protein-A affinity chromatography procedure. The overall yield for the downstream purification process is approximately 78%. The purifies of the purified mAb lots were around 94 - 99%, as determined by an analytical gel permeation chromatographic method. The purified mAb was active as reference lots of the antibody produced elsewhere, as determined by direct enzyme-linked immunosorbent assays.