Assessment of viral replication in eggs and HA protein yield of pre-pandemic H5N1 candidate vaccine viruses

Jing, Xiang-Hong; Soto, Jackeline; Gao, Yamei; Phy, Kathryn; Ye, Zhiping

doi:10.1016/j.vaccine.2013.07.011

Cited by 7 publications

(8 citation statements)

References 12 publications

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“…Bromelain‐cleaved HA was prepared essentially as described previously . Briefly, viruses were grown in 100 embryonated chicken eggs at 33°C by inoculation of 0·2 ml of diluted virus stock containing ~10 4 pfu and purified.…”

Section: Methodsmentioning

confidence: 99%

“…Bromelain-cleaved HA was prepared essentially as described previously. 10 Briefly, viruses were grown in 100 embryonated chicken eggs at 33°C by inoculation of 0Á2 ml of diluted virus stock containing~10 4 pfu and purified. Purified virus was diluted to a concentration of 10 mg/ml and incubated with bromelain, followed by br-HA purification through a continuous sucrose gradient.…”

Section: Preparation Of Bromelain-cleaved Ha and Vlps For Immunizationmentioning

confidence: 99%

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A novel approach for preparation of the antisera reagent for potency determination of inactivated H7N9 influenza vaccines

Schmeisser

Jing

Joshi

et al. 2016

Influenza Resp Viruses

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BackgroundThe potency of inactivated influenza vaccines is determined using a single‐radial immunodiffusion (SRID) assay and requires standardized reagents consisting of a Reference Antigen and an influenza strain‐specific antiserum. Timely availability of reagents is a critical step in influenza vaccine production, and the need for backup approaches for reagent preparation is an important component of pandemic preparedness.ObjectivesWhen novel H7N9 viruses emerged in China in 2013, candidate inactivated H7N9 influenza vaccines were developed for evaluation in clinical trials, and reagents were needed to measure vaccine potency.MethodsWe previously described an alternative approach for generating strain‐specific potency antisera, utilizing modified vaccinia virus Ankara vectors to produce influenza hemagglutinin (HA)‐containing virus‐like particles (VLPs) for immunization. Vector‐produced HA antigen is not dependent upon the success of the traditional bromelain‐digestion and HA purification.ResultsAntiserum for H7N9 vaccines, produced after immunization of sheep with preparations of bromelain‐HA (br‐HA), was not optimal for the SRID assay, and the supply of antiserum was limited. However, antiserum obtained from sheep boosted with VLPs containing H7 HA greatly improved the ring quality in the SRID assay. Importantly, this antiserum worked well with both egg‐ and cell‐derived antigen and was distributed to vaccine manufacturers.ConclusionsUtilizing a previously developed approach for preparing vaccine potency antiserum, we have addressed a major bottleneck encountered in preparation of H7N9 vaccine reagents. The combination of br‐HA and mammalian VLPs for sequential immunization represents the first use of an alternative approach for producing an influenza vaccine potency antiserum.

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Section: Methodsmentioning

confidence: 99%

Section: Preparation Of Bromelain-cleaved Ha and Vlps For Immunizationmentioning

confidence: 99%

A novel approach for preparation of the antisera reagent for potency determination of inactivated H7N9 influenza vaccines

Schmeisser

Jing

Joshi

et al. 2016

Influenza Resp Viruses

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“…16 The PLS was analyzed by deglycosylation treatment using a modified method based on previous reports. [17][18][19] Approximately 5 μg of viral protein was deglycosylated using PNGase F (New England Biolabs). The deglycosylated samples were separated by 10% SDS-PAGE and stained using a Colloidal Blue Staining Kit (Invitrogen).…”

Section: Calibration Of the Plsmentioning

confidence: 99%

“…The preparation method was modified based on a previous report. 18 Cell-derived H7N9 influenza virus was mixed with bromelain enzyme at a concentration of 0.1 U/mL (Sigma-Aldrich) containing β-mercaptoethanol (1:1000, v/v) and reacted for 3 h at RT with gentle shaking. The subsequent steps used for BHA preparation were identical to the steps used for XHA protein purification and SRID confirmation.…”

Section: Preparation Of Bha Protein From H7n9 Influenza Virusmentioning

confidence: 99%

Development and characterization of standard reagents for cell-based prepandemic influenza vaccine products

Lai

Weng

Chen

et al. 2020

Human Vaccines & Immunotherapeutics

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“…In some cases, more than two segments of the circulating (donor) strain are incorporated in the PR8-based IVS. IVSs suitable for large-scale manufacturing are required to replicate efficiently and to produce high amounts of HA per egg and/or viral particles with high HA content (Jing et al, 2013). As an alternative approach to embryonated eggs, specific mammalian cell lines that have been licensed for vaccine production by regulatory agencies (e.g.…”

Section: Introductionmentioning

confidence: 99%

The PB1 segment of an influenza A virus H1N1 2009pdm isolate enhances the replication efficiency of specific influenza vaccine strains in cell culture and embryonated eggs

Mostafa

Kanrai

Ziebuhr

et al. 2016

Journal of General Virology

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Influenza vaccine strains (IVSs) contain the haemagglutinin (HA) and neuraminidase (NA) genome segments of relevant circulating strains in the genetic background of influenza A/ PR/8/1934 virus (PR8). Previous work has shown that the nature of the PB1 segment may be a limiting factor for the efficient production of IVSs. Here, we showed that the PB1 segment (PB1 Gi ) from the 2009 pandemic influenza A virus (IAV) A/Giessen/06/2009 (Gi wt, H1N1pdm) may help to resolve (some of) these limitations. We produced a set of recombinant PR8-derived viruses that contained (i) the HA and NA segments from representative IAV strains (H3N2, H5N1, H7N9, H9N2); (ii) the PB1 segment from PR8 or Gi wt, respectively; and (iii) the remaining five genome segments from PR8. Viruses containing the PB1 Gi segment, together with the heterologous HA/NA segments and five PR8 segments (5+2+1), replicated to higher titres compared with their 6+2 counterparts containing six PR8 segments and the equivalent heterologous HA/NA segments. Compared with PB1 PR8 -containing IVSs, viruses with the PB1 Gi segment replicated to higher or similar titres in both cell culture and embryonated eggs, most profoundly IVSs of the H5N1 and H7N9 subtype, which are known to grow poorly in these systems. IVSs containing either the PB1 Gi or the cognate PB1 segment of the respective specific HA/NA donor strain showed enhanced or similar virus replication levels. This study suggests that substitution of PB1 PR8 with the PB1 Gi segment may greatly improve the large-scale production of PR8-derived IVSs, especially of those known to replicate poorly in vitro. INTRODUCTIONInfluenza viruses belong to the family Orthomyxoviridae and are classified into three genera termed Influenza virus A, B and C. Among these, influenza A viruses (IAVs) pose the greatest threat to human and animal health. They contain eight segments of negative-sense ssRNA (Ritchey et al., 1976;Shaw & Palese, 2013). The eight genomic segments encode at least 10 viral proteins, comprising the three subunits of the viral RNA polymerase complex [polymerase basic protein 2 (PB2), polymerase basic protein 1 (PB1) and polymerase acidic protein (PA)], the surface glycoproteins [haemagglutinin (HA) and neuraminidase (NA)], the nucleoprotein (NP), the matrix protein (M1), the ion channel protein (M2), the non-structural protein 1 (NS1) and the nuclear export protein (NEP). IAVs are genetically diverse, with new strains rapidly emerging by point mutations introduced by the low-fidelity viral RNA replicase (potentially resulting in antigenic drift if changes occur in the HA and/or NA coding sequences and affect important antigenic epitopes) or by genetic reassortment of genomic segments (potentially leading to antigenic shift if antigenically distant HA and NA segments are incorporated). Minor and major antigenic changes in the HA and NA proteins of circulating IAV are important driving forces for the emergence of new strains causing epidemics or even major pandemics (Neumann et al., 2009;Peng et al., 2014;Smith...

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Assessment of viral replication in eggs and HA protein yield of pre-pandemic H5N1 candidate vaccine viruses

Cited by 7 publications

References 12 publications

A novel approach for preparation of the antisera reagent for potency determination of inactivated H7N9 influenza vaccines

A novel approach for preparation of the antisera reagent for potency determination of inactivated H7N9 influenza vaccines

Development and characterization of standard reagents for cell-based prepandemic influenza vaccine products

The PB1 segment of an influenza A virus H1N1 2009pdm isolate enhances the replication efficiency of specific influenza vaccine strains in cell culture and embryonated eggs

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