Clinical and Diagnostic Laboratory Immunology, Jan 1994, 78-82, Vol 1, No. 1
Copyright © 1994 by the American Society for Microbiology. All rights reserved.

Enzyme-linked immunosorbent assay using a recombinant baculovirus- expressed Bacillus anthracis protective antigen (PA): measurement of human anti-PA antibodies

LC Iacono-Connors, J Novak, C Rossi, J Mangiafico and T Ksiazek
Virology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 21702-5011, USA.

We developed an antigen capture enzyme-linked immunosorbent assay (ELISA) which does not require purified protective antigen (PA) for detection of human antibodies to Bacillus anthracis PA. Lysates of Spodoptera frugiperda (Sf-9) cells infected with recombinant baculovirus containing the PA gene were used as the source of PA to develop the ELISA. Recombinant PA from crude Sf-9 cell lysates or PA purified from B. anthracis Sterne strain was captured by an anti-PA monoclonal antibody coated onto microtiter plates. We demonstrated that human serum antibody titers to PA were identical in the ELISA whether we used crude Sf-9 cell lysates containing recombinant baculovirus- expressed PA or purified Sterne PA. Finally, false-positive results observed in a direct ELISA were eliminated with this antigen capture ELISA. Thus, the antigen capture ELISA with crude preparations of baculovirus-expressed PA is reliable, safe, and inexpensive for determining anti-PA antibody levels in human sera.

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