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

Abstract

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.

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Document Details

Document Type
Technical Report
Publication Date
Jun 01, 2002
Accession Number
ADA405736

Entities

People

  • Cheng J. Cao
  • Derrel Menking
  • Jun T. Park
  • Sarah J. Cork
  • Tracy Collano

Organizations

  • Edgewood Chemical Biological Center

Tags

Communities of Interest

  • Human Systems

DTIC Thesaurus Topics

  • Albumins
  • Anti-Bacterial Agents
  • Biological Warfare
  • Biological Warfare Agents
  • Biosensors
  • Cell Line
  • Cells
  • Chromatography
  • Culture Media
  • Culture Techniques
  • Detection
  • Detectors
  • Gammaproteobacteria
  • Liquid Chromatography
  • Production
  • Proteins
  • Sodium Compounds

Fields of Study

  • Biology

Readers

  • Analytical Chemistry
  • Immunology

Technology Areas

  • Biotechnology