Method of making a vaccine for anthrax

Details Method of making a vaccine for anthrax Method of making a vaccine for anthrax

2000-03-07

2002-05-14

Graser, Jennifer E. (Department: 1643)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Using a micro-organism to make a protein or polypeptide

C435S069100, C435S069400, C435S320100, C435S173300, C435S173300, C435S252300, C435S200000, C435S252310, C530S350000, C424S184100, C424S234100, C424S246100

Reexamination Certificate

active

06387665

ABSTRACT:

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. [Iacono-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.


REFERENCES:
patent: 3208909 (1965-09-01), Puziss et al.
patent: 4455142 (1984-06-01), Martins et al.
patent: 5071748 (1991-12-01), Miller
patent: 5077214 (1991-12-01), Guarino et al.
patent: 5081029 (1992-01-01), Zarling et al.
Ivins et al. European J. Epidemiology. Mar. 1988. 4(1): 12-19*
Ivins et al. Clin. Immunology Newsletter. 9(2): 30-32, 1988.*
Uptake of Congo red by Virulent Strains of Bacillus Anthracis, Worsham et al., 1991 ASM Abstracts, p. 75.
Understanding Biotechnology Law, edited by Gale R. Peterson, Marcel Dekker, Inc., (date unknown).
Vaccine Efficacy of Bacillus anthracis Protective Antigen Produced in Pokaryotic and Eukaryotic Cells, Ivins, et al., Inst. Infect. Dis., Ft. Detric Frederick, MD, May 25, 1994.
Immunization Against Anthrax with Aromatic Compound Dependent (Aro-) Mutants of Bacillus anthracis and with Recombinant Strains of Bacillus subtilis that Produce Anthrax Protective Antigen; Ivins, et al., Infect. and Immunity, Nov. 1986, vol. 54, No. 2, pp. 537-542.
Cloning and Expression of teh Bacillus anthracis, Ivins, et al.; Infect and Immunity, May 1986, vol. 52, No. 2, pp. 454-458.
Expression of the Bacillus antracis Protective Antigen Gene by Baculovirus and Vaccinia Virus Recombinants; Icono-Connors, et al., Infect. and Immunity, Feb. 1990, vol. 58, No. 2, pp. 366-372.
Protection against Anthrax with Recombinant Virus-Expressed Protective Antigen in Experimental Animals; Iacono-Connors, et al., Infect. and Immunity, Jun. 1991, vol. 59, No. 6, pp. 1961-1965.

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