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Amerithrax — Part 27
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United States Patent: 6,387,665 e @ Page 3 of 11
3. The method of claim 1, wherein the vaccine is in the form of a suspension.
4. The method of claim 1 wherein the vaccine is in the form of buffered suspension.
5. The method of claim 1 wherein said carrier is an adjuvant.
Description
FIELD OF THE INVENTION
This invention relates to the bacterial expression system, production and use of protective antigen (PA)
against Bacillus anthracis. The PA immunogen is useful in vaccine against human anthrax. The PA can
be produced by an asporogenic organism which overproduces the desired antigen, which is then
harvested from the supernatant.
BACKGROUND OF THE INVENTION
Bacillus anthracis is the etiologic agent responsible for anthrax, a disease often found in persons
exposed to infected animals or their products. Persons particularly exposed to animals include
veterinarians, laboratory technicians, ranchers and employees working with skin or hair of animals. The
mode of entry into the body may be the skin or, when contaminated meat is eaten, the gastrointestinal
tract. Inhaling of spores can cause inhalation anthrax, a disease that can be fatal. Vaccines against
Bacillus anthracis have been available. Virulent strains of the organism produce two toxins and a poly-
D-glutamic acid capsule which are coded for on two endogenous plasmids, pX01 and pX02,
respectively. Loss of either of the plasmids results in an attenuated strain of reduced virulence, while
loss of both results in an avirulent organism. The history of the USAMRIID Sterne strain of B. anthracis
prior to 1981 is uncertain, though it is believed to be derived from the Sterne strain isolated at the
Onderstpoort Research Laboratory in Pretoria, South Africa.
In 1985 the Bacillus anthracis protective antigen (PA) gene was cloned into a plasmid (pUB110)
resulting in the formation of a recombinant plasmid identified as pPA102, which was reported in the
literature (Ivins and Welkos, Infection and Immunity, 54:537-542 (1986)). The production of vaccines
lacking lethal factor was possible thereby. However, a primary problem remained, since the Bacillus
anthracis formed spores. Once spores have formed, they persist in the environment for months and
years. Once the laboratory environment contains such spores, it is very difficult to free the environment
of the spores.
It was also previously reported that protective antigen (PA) could be produced in baculovirus. [Jacono-
Connors, et al., Infection and Immunity, 58:366-372 (1990); Iacono-Connors, et al., Infection and
Immunity, 59:1961-1965 (1991)] A major problem in production of the PA in the baculovirus disclosed
therein is that the desired antigen requires a complex purification process. Even after purification by
immuno-affinity chromatography, undesired cellular material continues to contaminate the desired
product.
DETAILED DESCRIPTION OF THE INVENTION
The instant invention provides organisms which produce protective antigen (PA) lacking lethal factor
and edema factor proteins which, when present as contaminants in vaccine, can cause serious side
http://patft.uspto.gov/netacgi/nph-Parser?Sectl=PTO2&Sect2=HITOFF &u=/netahtml/sear... 6/28/2005
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