Biophysical characterization and immunization studies of dominant negative inhibitor (DNI), a candidate anthrax toxin subunit vaccine.
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.
Macromolecule and Vaccine Stabilization Center; Department of Pharmaceutical Chemistry; University of Kansas; Lawrence, KS USA; Bioengineering Graduate Program; University of Kansas; Lawrence, KS USA.
Vidyashankara
V
Iyer
Lei
L
Hu
Carole E
CE
Schanté
David
D
Vance
Chrystal
C
Chadwick
Nishant Kumar
NK
Jain
Robert N
RN
Brey
Sangeeta B
SB
Joshi
David B
DB
Volkin
Kiran K
KK
Andra
James G
JG
Bann
Nicholas J
NJ
Mantis
C Russell
CR
Middaugh
D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000536:Aluminum Hydroxide / Q000008:administration & dosage; D000818:Animals; D000881:Anthrax / Q000517:prevention & control*; D022122:Anthrax Vaccines / Q000008:administration & dosage / Q000276:immunology*; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000276:immunology*; D001408:Bacillus anthracis / Q000276:immunology*; D001427:Bacterial Toxins / Q000737:chemistry* / Q000276:immunology*; D055592:Biophysical Phenomena; D004355:Drug Stability; D005612:Freeze Drying; D007114:Immunization / Q000379:methods*; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D055550:Protein Stability / Q000528:radiation effects; D021122:Protein Subunits / Q000737:chemistry / Q000276:immunology; D013696:Temperature; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000276:immunology
D000276:Adjuvants, Immunologic; D022122:Anthrax Vaccines; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D021122:Protein Subunits; D022223:Vaccines, Subunit; C030325:anthrax toxin; D000536:Aluminum Hydroxide
Accelerated Stability; Aluminum Hydroxide; Biophysical characterization; Dominant-Negative Inhibitor; Immunogenicity; Vaccine