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Hum Vaccin Immunother. 2013 Nov;9(11):2362-70. Epub 2013 Aug 7.

Biophysical characterization and immunization studies of dominant negative inhibitor (DNI), a candidate anthrax toxin subunit vaccine.

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Macromolecule and Vaccine Stabilization Center; Department of Pharmaceutical Chemistry; University of Kansas; Lawrence, KS USA; Bioengineering Graduate Program; University of Kansas; Lawrence, KS USA.


Dominant Negative Inhibitor (DNI) is a translocation-deficient homolog of recombinant protective antigen of Bacillus anthracis that is a candidate for a next generation anthrax vaccine. This study demonstrates that the biophysical characteristics of the DNI protein stored in lyophilized form at 4°C for 8 y were similar to recombinant Protective Antigen (rPA). To provide information on the accelerated stability of DNI, samples in the lyophilized form were subjected to thermal stress (40°C and 70°C for up to 4 weeks) and thoroughly evaluated using various biophysical and chemical characterization techniques. Results demonstrate preserved structural stability of the DNI protein under extreme conditions, suggesting long-term stability can be achieved for a vaccine that employs DNI, as desired for a biodefense countermeasure. Furthermore, the biological activity of the stressed DNI bound to the adjuvant Alhydrogel (®) was evaluated in mice and it was found that the immunogenicity DNI was not affected by thermal stress.


Accelerated Stability; Aluminum Hydroxide; Biophysical characterization; Dominant-Negative Inhibitor; Immunogenicity; Vaccine

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