H1N1 Influenza Virus-Like Particles: Physical Degradation Pathways and Identification of Stabilizers

Kissmann, Julian; Joshi, Sangeeta B.; Haynes, Joel R.; Dokken, Leslie; Richardson, Charles C.; Middaugh, C. Russell

doi:10.1002/jps.22304

Cited by 25 publications

(18 citation statements)

References 32 publications

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“…Radar chart analysis is a data visualization tool that has been used to summarize and compare large biophysical data sets to study the effect of various stresses (e.g., pH, temperature) on physical stability of a wide variety of macromolecules including vaccine and protein drug candidates. 34,[51][52][53][54][55][56][57] Radar chart analysis in this work showed the structural integrity and conformational stability of 3 NRRV antigens as a function of pH and temperature. P [8] was the most stable, followed by P [4], and P [6] was least stable.…”

Section: Size Higher-order Structure and Physical Stability Profilementioning

confidence: 99%

Recombinant Subunit Rotavirus Trivalent Vaccine Candidate: Physicochemical Comparisons and Stability Evaluations of Three Protein Antigens

Hickey

Sahni

Toth

et al. 2020

Journal of Pharmaceutical Sciences

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a b s t r a c tAlthough live attenuated Rotavirus (RV) vaccines are available globally to provide protection against enteric RV disease, efficacy is substantially lower in low-to middle-income settings leading to interest in alternative vaccines. One promising candidate is a trivalent nonreplicating RV vaccine, comprising 3 truncated RV VP8 subunit proteins fused to the P2 CD4 þ epitope from tetanus toxin (P2-VP8-P[4/6/8]). A wide variety of analytical techniques were used to compare the physicochemical properties of these 3 recombinant fusion proteins. Various environmental stresses were used to evaluate antigen stability and elucidate degradation pathways. P2-VP8-P[4] and P2-VP8-P[6] displayed similar physical stability profiles as function of pH and temperature while P2-VP8-P[8] was relatively more stable. Forced degradation studies revealed similar chemical stability profiles with Met 1 most susceptible to oxidation, the single Cys residue (at position 173/172) forming intermolecular disulfide bonds (P2-VP8-P[6] was most susceptible), and Asn 7 undergoing the highest levels of deamidation. These results are visualized in a structural model of the nonreplicating RV antigens. The establishment of key structural attributes of each antigen, along with corresponding stability-indicating methods, have been applied to vaccine formulation development efforts (see companion paper), and will be utilized in future analytical comparability assessments.

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Section: Size Higher-order Structure and Physical Stability Profilementioning

confidence: 99%

Recombinant Subunit Rotavirus Trivalent Vaccine Candidate: Physicochemical Comparisons and Stability Evaluations of Three Protein Antigens

Hickey

Sahni

Toth

et al. 2020

Journal of Pharmaceutical Sciences

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“…Similar results were obtained with VLPs containing the minigenome with the weak S segment promoter (data not shown). We also tested the influence of various additives known to enhance VLP stability (28,29) (Fig. 4D).…”

Section: Generation Of Cchf Virus-like Particles Expressing a Reportementioning

confidence: 99%

A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

Devignot

Bergeron

Nichol

et al. 2015

J Virol

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Crimean-Congo hemorrhagic fever virus (CCHFV; genus Nairovirus) is an extremely pathogenic member of the Bunyaviridae family. Since handling of the virus requires a biosafety level 4 (BSL-4) facility, little is known about pathomechanisms and host interactions. Here, we describe the establishment of a transcriptionally competent virus-like particle (tc- VLP IMPORTANCECrimean-Congo hemorrhagic fever virus (CCHFV) is an extremely virulent pathogen of humans. Since the virus can be handled only at the highest biosafety level, research is restricted to a few specialized laboratories. We developed a plasmid-based system to produce virus-like particles with the ability to infect cells and transcribe a reporter genome. Due to the absence of viral genes, the virus-like particles are unable to spread or cause disease, thus allowing study of aspects of CCHFV biology under relaxed biosafety conditions. C rimean-Congo hemorrhagic fever (CCHF) is a severe viral disease in Eastern Europe, the Middle East, Asia, and Africa, reported to have a case/fatality rate of approximately 30% (1). Infections of humans are associated with an acute febrile disease that can lead to hemorrhages, hypovolemic shock, and death, while in infected animals no clinical signs can be detected. The CCHF virus (CCHFV) is transmitted via tick bites (principally from the Hyalomma genus) or by direct contact with blood or tissues from infected persons or animals (2). The efficiency of ribavirin as an antiviral in humans is still under debate (3), and there are currently no established prophylaxis, no specific treatment, and no FDA-approved vaccine. The pathogenesis as well as immune responses are poorly characterized, mostly due to the restriction to high-biosafety-level facilities (biosafety level 4 [BSL-4]) for handling of virus and biological samples. The only animal models available so far are based on mice lacking antiviral interferon (IFN) responses, thus hampering studies on innate immune system interactions (4, 5). Clearly, there is paucity in tools and methods to better study the virus and its host cell interactions.CCHFV belongs to the Nairovirus genus of the family Bunyaviridae. This enveloped virus has a single-stranded negative-sense RNA genome (viral RNA [vRNA]) of an unusually large total size of 19,146 nucleotides (nt). The genome is divided into 3 segments and encodes 4 structural proteins: the RNA-dependent RNA polymerase (RdRp) L on the large (L) segment (12,108 nt), the two glycoproteins (GPs) Gc and Gn on the medium (M) segment (5,366 nt), and the nucleoprotein N on the small (S) segment (1,672 nt). The nucleoprotein N binds each RNA segment to form ribonucleoprotein complexes (RNPs), the functional template of the viral polymerase (pol) L for transcription and replication. Shortly after infection, CCHFV performs the so-called primary transcription leading to initial mRNA synthesis. The protein products are necessary for subsequent replication of the viral genome, resulting in copy RNA (cRNA) (positive sense) and progeny genom...

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“…EPDs provide a convenient effective way to summarize large amounts of complementary data and provide a comprehensive view of VLP structural changes induced by change in environmental factors. 40,41 Because of the gradual signal changes observed in many of the techniques utilized in the study, especially for samples around neutral pH, there was some difficulty in defining the apparent phase boundaries to delineate the clear structural changes of eVLPs and mVLPs under neutral pH conditions. In the case of eVLPs, there are approximately four different phases existing at low temperature (Fig.…”

Section: Empirical Phase Diagramsmentioning

confidence: 99%

Biophysical Characterization and Conformational Stability of Ebola and Marburg Virus-Like Particles

Trefethen

Zeng

et al. 2011

Journal of Pharmaceutical Sciences

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H1N1 Influenza Virus-Like Particles: Physical Degradation Pathways and Identification of Stabilizers

Cited by 25 publications

References 32 publications

Recombinant Subunit Rotavirus Trivalent Vaccine Candidate: Physicochemical Comparisons and Stability Evaluations of Three Protein Antigens

Recombinant Subunit Rotavirus Trivalent Vaccine Candidate: Physicochemical Comparisons and Stability Evaluations of Three Protein Antigens

A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

Biophysical Characterization and Conformational Stability of Ebola and Marburg Virus-Like Particles

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