Physical Characterization of Clostridium Difficile Toxins and Toxoids: Effect of the Formaldehyde Crosslinking on Thermal Stability

Salnikova, Maya S.; Joshi, Sangeeta B.; Rytting, J. Howard; Warny, Michel; Middaugh, C. Russell

doi:10.1002/jps.21261

Cited by 37 publications

(44 citation statements)

References 43 publications

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“…Although a number of C. difficile vaccines are being developed, their utility has been limited by the need for repetitive dosing, the use of formalin-induced detoxification, and the stability of the vaccine constructs (30,31). For a vaccine to warrant clinical development it must induce a strong protective immune response against autologous and heterologous strains, requiring the fewest immunizations with or without the added help of an adjuvant.…”

Section: Discussionmentioning

confidence: 99%

Toll-Like Receptor 5-Dependent Immunogenicity and Protective Efficacy of a Recombinant Fusion Protein Vaccine Containing the Nontoxic Domains of Clostridium difficile Toxins A and B and Salmonella enterica Serovar Typhimurium Flagellin in a Mouse Model of Clostridium difficile Disease

et al. 2013

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Section: Discussionmentioning

confidence: 99%

Toll-Like Receptor 5-Dependent Immunogenicity and Protective Efficacy of a Recombinant Fusion Protein Vaccine Containing the Nontoxic Domains of Clostridium difficile Toxins A and B and Salmonella enterica Serovar Typhimurium Flagellin in a Mouse Model of Clostridium difficile Disease

et al. 2013

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“…The large data sets, obtained from techniques such as circular dichroism, intrinsic and extrinsic fluorescence and light scattering, have been used previously to evaluate vaccine stability and were used to establish a stability map ("EPD") of Na-GST-1. [21][22][23][24][25][26][27][28] Circular dichroism studies. Changes in the secondary structure of Na-GST-1 were studied by monitoring the change in the far UV-CD signal as a function of both pH and temperature.…”

Section: Biophysical Characterization and Construction Of A Multiple mentioning

confidence: 99%

Advances in vaccines against neglected tropical diseases

Plieskatt

Rezende

Olsen

et al. 2012

Human Vaccines & Immunotherapeutics

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A bivalent recombinant vaccine for human hookworm disease is under development. One of the lead candidate antigens in the vaccine is a glutathione S-transferase cloned from the hookworm Necator americanus (Na-GST-1) which is expressed in the yeast Pichia pastoris. Based on preliminary studies demonstrating that the recombinant protein was not stable in an acetate buffer at pH 6, we undertook an extensive stability analysis of the molecule. To improve and optimize stability we complemented traditional methods employed for macromolecule and vaccine stabilization with biophysical techniques that were incorporated into a systematic process based on an eigenvector approach. Large data sets, obtained from a variety of experimental methods were used to establish a color map ("empirical phase diagram") of the physical stability of the vaccine antigen over a wide range of temperature and pH. The resulting map defined "apparent phase boundaries" that were used to develop high throughput screening assays. These assays were then employed to identify excipients that stabilized the antigen against physical degradation that could otherwise result in losses of physicochemical integrity, immunogenicity, and potency of the vaccine. Based on these evaluations, the recombinant Na-GST-1 antigen was reformulated and ultimately produced under Good Manufacturing Practices and with an acceptable stability profile.

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“…This method has been successfully applied previously for the characterization of different proteins 31 including mAbs, 14 as well as larger macromolecular complexes such as viruses, 32 toxins and toxoids, 33 and various vaccines. 34 Eigenvectors of the density matrix were obtained using the data sets from the various biophysical techniques described above to generate color maps (EPDs) of the proteins' conformational behavior as a function of pH and temperature.…”

Section: Empirical Phase Diagramsmentioning

confidence: 99%

Formulation Design and High-Throughput Excipient Selection Based on Structural Integrity and Conformational Stability of Dilute and Highly Concentrated IgG1 Monoclonal Antibody Solutions

Bhambhani

Kissmann

Joshi

et al. 2012

Journal of Pharmaceutical Sciences

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Physical Characterization of Clostridium Difficile Toxins and Toxoids: Effect of the Formaldehyde Crosslinking on Thermal Stability

Cited by 37 publications

References 43 publications

Toll-Like Receptor 5-Dependent Immunogenicity and Protective Efficacy of a Recombinant Fusion Protein Vaccine Containing the Nontoxic Domains of Clostridium difficile Toxins A and B and Salmonella enterica Serovar Typhimurium Flagellin in a Mouse Model of Clostridium difficile Disease

Toll-Like Receptor 5-Dependent Immunogenicity and Protective Efficacy of a Recombinant Fusion Protein Vaccine Containing the Nontoxic Domains of Clostridium difficile Toxins A and B and Salmonella enterica Serovar Typhimurium Flagellin in a Mouse Model of Clostridium difficile Disease

Advances in vaccines against neglected tropical diseases

Formulation Design and High-Throughput Excipient Selection Based on Structural Integrity and Conformational Stability of Dilute and Highly Concentrated IgG1 Monoclonal Antibody Solutions

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