Production and Validation of an IgM Antibody for the Detection of Spore-Forming Bioagents
Final rept. May 2002-May 2003
EDGEWOOD CHEMICAL BIOLOGICAL CENTER ABERDEEN PROVING GROUND MD RESEARCH AND TECHNOLOGY DIR
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Antibodies are the essential components in biosensors that specifically bind biological agents, providing a capability essential to fulfilling the defense mission of Department of Defense. An IgM monoclonal antibody specific for the spore forming Bacillus anthracis has been developed by the John Kearney Laboratory University of Alabama, Birmingham, AL. However, a means of purifying IgM in quantities sufficient for testing and evaluation was not readily available. The IgM has a monomeric subunit structure consisting of two heavy and two light chains. However, IgM is efficiently secreted only if it is polymerized into either pentamers or hexamers, 750-900K Daltons. Unlike IgG, IgM is not effectively purified by either protein-A or protein-G. These properties introduce difficulties in the IgM antibody purification. In this study, a platform process was developed for the purification of IgM, including an immuno-affinity chromatography method using a goat anti-mouse IgM antibody coupled to Sepharose as a polishing chromatographic step. The purified IgM was active as determined by ELISA.
- Chemical, Biological and Radiological Warfare
- Physical Chemistry