Treatment of experimental anthrax with pegylated circularly permuted capsule depolymerase

Legler, Patricia M.; Little, Stephen F.; Senft, Jeffrey; Schokman, Rowena D; Carra, John H.; Compton, Jaimee R.; Chabot, Donald J.; Tobery, Steven A.; Fetterer, David P.; Siegel, Justin B.; Baker, David; Friedlander, Arthur M.

doi:10.1126/scitranslmed.abh1682

Cited by 2 publications

(10 citation statements)

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“…50 The data on Adagen provided some useful insight regarding the safety, efficacy, and immunogenicity of pegylated biologics. For PEG-CapD S334C , the treatment period that we previously tested was 2 days with a total of 6 injections occurring every 8 h (40 mg/kg, 80% survival), 13 a relatively short treatment period when compared with antibiotics such as ciprofloxacin. Acute bacterial infections, unlike SCID (a chronic condition), require shortterm treatment.…”

Section: ■ Discussionmentioning

confidence: 99%

“…In our previous study, 13 we examined a 3-prong branched PEG-CapD S334C -CP with a TEV protease cleavage site. The linear mPEG-CapD-CP gave an AUC that was significantly higher than what we obtained for the branched PEG-CapD S334C -CP after a 40 mg/kg dose.…”

Section: Enhancement Of Avidity For the High Molecular Weight Capsula...mentioning

confidence: 99%

“…In bacteria, the fmethionine is removed from the N-terminus, and the activity of the purified CapD-CP is ∼2-fold higher than that of the natural CapD. In our recent work, 23 we found that the pegylated form of CapD-CP (PEG-CapD-CP S334C ) was effective in protecting mice from lethal challenges of ΔAmes (an encapsulated strain lacking the pXO1 plasmid) and Ames spores. The enzyme was delivered 24 h postinfection every 8 h for 2 days at a concentration of 40 mg/kg.…”

mentioning

confidence: 99%

“…We developed a pegylated CapD S334C -CP enzyme, an Fc-fusion of the CapD S334C -CP enzyme, and a pegylated Fc-fusion (PEG-Fc-CapD S334C -CP). In our previous work, 13 we tested a 3-prong branched TMM(PEG) 12 molecule on a CapD-CP construct with a TEV protease cleavage site and His-tag (Figure S2).…”

mentioning

confidence: 99%

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Engineering an Fc-Fusion of a Capsule Degrading Enzyme for the Treatment of Anthrax

Matharoo

Chua

Park³

et al. 2022

ACS Infect. Dis.

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Polymers of d-glutamic acid (PDGA) form the capsule of the highly virulent Ames strain of B. anthracis. PDGA is antiphagocytic and weakly immunogenic; it enables the bacteria to evade the innate immune responses. CapD is an enzyme that catalyzes the covalent anchoring of PDGA. CapD is an Ntn-amido hydrolase that utilizes an internal Thr-352 as its nucleophile and general acid and base. An internal cleavage produces a free N-terminal Thr-352 and a short and long polypeptide chain. The chains were circularly permuted (CP) to move Thr-352 to the N-terminus of the polypeptide. We previously showed that a branched PEG-CapDS334C-CP could protect mice (80% survival) against a 5 LD50 challenge with B. anthracis Ames without the use of antibiotics, monoclonals, or vaccines. In attempts to improve the in vivo circulation time of CapD and enhance its avidity to its polymeric substrate, an Fc-domain of a mouse IgG1 was fused to CapDS334C-CP and the linker length and sequence were optimized. The resulting construct, Fc-CapDS334C-CP, then was pegylated with a linear 2 kDa mPEG at S334C to produce mPEG-Fc-CapDS334C-CP. Interestingly, the fusion of the Fc-domain and incorporation of the S334C mutation imparted acid stability, but slightly reduced the k cat (∼ 2-fold lower). In vivo, the measured protein concentration in sera was higher for the Fc-fusion constructs compared to the mPEG-Fc-CapDS334C-CP. However, the exposure calculated from measured sera enzymatic activity was higher for the mPEG-CapDS334C-CP. The pegylated Fc-fusion was less active than the PEG-CapDS334C-CP, but detectable in sera at 24 h by immunoblot. Here we describe the engineering of a soluble, active, pegylated Fc-fusion of B. anthracis CapD (mPEG-Fc-CapD-CP) with activity in vitro, in serum, and on encapsulated bacteria.

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Section: ■ Discussionmentioning

confidence: 99%

Section: Enhancement Of Avidity For the High Molecular Weight Capsula...mentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Engineering an Fc-Fusion of a Capsule Degrading Enzyme for the Treatment of Anthrax

Matharoo

Chua

Park³

et al. 2022

ACS Infect. Dis.

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“…Those vaccines are called subunit vaccines and have significant advantages over traditional vaccines that include improved immunity (if the suitable antigen is selected), specificity, compatibility, accessible storage and maintenance (12). However, that leads to increased demand for determining the appropriate formulation to produce an immune response (13).…”

Section: Introductionmentioning

confidence: 99%

A recombinant chimeric protein of protective antigen and lethal factor from Bacillus anthracis in polymeric nanocapsules showed a strong immune response in mice: a potential high efficacy vaccine against anthrax

Aziziaram¹,

Mattoo²

2022

Cell Mol Biol (Noisy-le-grand)

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Anthrax is a serious infectious disease caused by Bacillus anthracis, rod-shaped gram-positive bacteria. The disease infects both humans and animals and causes severe illness. Many vaccines have been developed for anthrax, but the vaccine with very high efficacy is yet to be developed. To overcome the problems of efficacy posed by the existing vaccines, a recombinant chimeric fusion protein containing domain 1 of lethal factor (LFD1) and domain 4 of Bacillus anthracis protective antigen (PA4) was used as antigen in copolymeric nanocapsules (NCs). Accordingly, the solvent evaporation double emulsion method was used to produce NCs containing recombinant chimeric fusion protein (LFD1-PA4). Zeta sizer and potential of nanoparticles, nanoparticle loading efficiency, release pattern of recombinant protein, and the possible effect of polylactic acid-polyethylene glycol (PLA-PEG) nanoparticle production method were investigated. Mice were used to test and evaluate the immune response. The mean titer of antibody produced against loaded LFD1-PA4 compared to free form showed a significant difference. The difference in antibody titer between the groups of once injected, twice injected, and free antigen was significant, and the highest antibody titer was found in the mice twice injected. In addition, a single-time loaded injection showed significantly higher antibodies than the free form injection indicating that loaded LFD1-PA4 into PLA-PEG nanoparticles elicits a stronger immune response. This study showed that LFD1-PA4 fusion protein from Bacillus anthracis served as an active antigen in mice. Also, the nanocarrier (PLA-PEG) containing the antigen can stimulate the immune system in the mice, owing to their controlled release property.

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Treatment of experimental anthrax with pegylated circularly permuted capsule depolymerase

Abstract: Pegylated circularly permuted capsule depolymerase removes capsule from Bacillus anthracis and protects anthrax-infected mice.

Cited by 2 publications

References 35 publications

Engineering an Fc-Fusion of a Capsule Degrading Enzyme for the Treatment of Anthrax

Engineering an Fc-Fusion of a Capsule Degrading Enzyme for the Treatment of Anthrax

A recombinant chimeric protein of protective antigen and lethal factor from Bacillus anthracis in polymeric nanocapsules showed a strong immune response in mice: a potential high efficacy vaccine against anthrax

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