Guidance for industry potency tests for cellular and gene therapy products. WHO Workshop in Training Performance of Potency Testing and Lot Release of Human Papillomavirus (HPV) Vaccines, 1115 May (2009). Stitz, L. et al. EMA. For example, for a recombinant protein antigen, dose (or content) measurement is typically made by a quantitative protein assay or near-ultraviolet (UV) absorbance, while potency is measured by an immunoassay, at a series of pre-determined doses, as a surrogate of biological immune response. 19, 667668 (2020). Rev. Development and licensure of vaccines to prevent COVID-19. authorstream presentation slide 17 rabies anti serum rabies anti serum is a preparation containing the Walsh, E. E. et al. EDQM. These measurements and quantitative determination of doses to be delivered must be as accurate and precise as possible, though the particular assays for quantifying these CQA depend on the platform and the product being administered to the vaccine recipients. . CEPI selected ELISA methods that exploit and capture signals of full-length SARS-CoV-2 S, RBD, and N-protein IgG antibody/antigen interactions. Vaccine Research and Development, Coalition for Epidemics Preparedness Innovations, CEPI, Torshov, Oslo, Norway, Gautam Sanyal,Anna Srneflt&Arun Kumar, You can also search for this author in This leads to several important differences in the characteristics of the resulting products. J. Virol. Safety and immunogenicity of a 2-dose heterologous vaccination regimen with Ad26.ZEBOV and MVA-BN-Filo Ebola vaccines: 12-month data from a phase 1 randomized clinical trial in Uganda and Tanzania. April 20th, 2019 - Rabies vaccine is a vaccine used to prevent rabies There are a number of vaccines available that are . People who have a history of serious hypersensitivity to components of rabies vaccine should be revaccinated with caution. Pain, redness, or swelling where the shot was given. & Hennessey, J. P. Evaluation of accuracy and precision of adenovirus absorptivity at 260 nm under conditions of complete DNA disruption. FDA Issues Emergency Use Authorization for Third COVID-19 Vaccine (2021). The rabies vaccine triggers your immune system to produce antibodies, Dodge explains. Guidance for Industry. Mild, local reactions to the rabies vaccine, such as pain, redness, swelling, or itching at the injection site, have been reported. QPA combines qPCR with a tissue culture-based infectivity assay to quantify the adenovirus (prime) potency, while the boost MVA-BN-Filo potency is determined solely on infectivity through a flow cytometry-based method73. Biotechnol. It took less than a year from the publication of SARS-CoV-2 gene sequence to Emergency Use Authorization (EUA) of the first vaccine, setting a record for speed in the history of vaccine development. Kuhn, I., Larsen, B., Gross, C. & Hermiston, T. High-performance liquid chromatography method for rapid assessment of viral particle number in crude adenoviral lysates of mixed serotype. The cost of the rabies vaccine continues to be a limitation to acquiring pre-exposure rabies immunization for travelers from developed countries. Because the virus . Phase 1 assessment of the safety and Immunogenicity of an mRNA- lipid nanoparticle vaccine candidate against SARS-CoV-2 in human volunteers. Drug Discov. https://doi.org/10.1371/journal.pone.0024135 (2011). The rabies. Analytical evaluation methods, that can ensure the high quality of the products and that can be executed speedily, must be in place as an integral component of Chemistry, Manufacturing, and Control (CMC). An mRNA vaccine encoding rabies virus glycoprotein induces protection against lethal infection in mice and correlates of protection in adult and newborn pigs. Demonstration of comparability between phase 3 and commercial lots is required and is especially critical if scale up or scale out is involved, even though drug substance (DS) and drug product (DP) processes were locked prior to phase 3. An in vitro potency assay for a sa-mRNA construct was developed to measure replication efficiency by capturing intermediate double-stranded RNA (dsRNA) and comparable protein expression in individual cells by an antigen-specific antibody. pDNA vaccines are best delivered by electroporation using a special device after intramuscular or subcutaneous injections. Badger, C. V. et al. In particular, we will focus on potency assays, which are key to delivering safe and immunogenic doses of vaccines. The authors thank Tung Thanh Le and COVID-19 vaccine landscape team of CEPI for providing critical and detailed input. https://www.gov.uk/government/publications/regulatory-approval-of-covid-19-vaccine-astrazeneca/conditions-of-authorisation-for-covid-19-vaccine-astrazeneca (2020). This is called post-exposure prophylaxis. Other stability indicating CQA includes physical, chemical, and structural integrity. and JavaScript. They are equally safe and effective. While clinical (human) efficacy of a vaccine can never be predicted from pre-clinical evaluation of CQA, precise and accurate measurements of antigen content and a relevant biological activity (termed potency) elicited by the antigen allow selection of potentially safe and immunogenic doses for entry into clinical trials. Another rapid analysis method for viral particle quantification is high-performance liquid chromatography (HPLC) where the column allows separation of intact virus particles from other cellular contaminants or fragmented virus particles76. 54, 6275 (2020). Interestingly, immunogenic profile of Zika virus sa-mRNA vaccine was found to be comparable with frequencies of dsRNA positive BHK cells and protein expression53. Assessment report Mvabea. Although assays are established for the proven platforms, such as LAV and recombinant proteins, faster and more robust in vitro assays can be developed for some CQA. Hussain, A. I., Cordeiro, M., Sevilla, E. & Liu, J. We discuss here opportunities to improve the speed and quality of the critical batch release and characterization assays. Vaccinating against rabies to save lives Human rabies is a 100% vaccine-preventable disease, yet it continues to kill. Rauch, S. et al. Biopharm. ISSN 2059-0105 (online). Russia files Sputnik V vaccine registration in Europe (2021). Vaccine 23, 45004508 (2005). Evidence for multiple protein components in the virion and a comparison of types 2, 7A and 12. Furthermore, CLs in CEPIs global network of laboratoriess are using common key reagents including coating protein, SARS-CoV-2 virus strain, virus pseudoparticles, peptide pools for PBMC stimulation, panel of standards and controls. Rose, Paul Stickings, Chris Burns, Rose T. Byrne-Nash, Jacob H. Gillis, Kathy L. Rowlen, Rose T. Byrne-Nash, David F. Miller, Kathy L. Rowlen, Natalie Thiel, Casey Selwyn, Ajoy C. Chakrabarti, Catherine Milne, Ralf Wagner, Laurent Mallet, Giuditta Guerrini, Davide Magr, Luigi Calzolai, Viki Bockstal, Auguste Gaddah, Macaya Douoguih, npj Vaccines In addition, the viral vector platform, which earlier yielded two vaccines against Ebola, using both replicating and non-replicating vectors5,6,7, has now delivered vaccines against SARS-CoV-28,9,10,11,12. Considerations for bioanalytical characterization and batch release of COVID-19 vaccines. Potency is also a primary stability indicator and will need to be monitored as a function of time for all candidate antigens in bulk (DS) as well as formulated DP. Maintaining comparability between smaller scale material such as that often used in pre-clinical toxicity study and phase 1 trial with later phase clinical trial material (CTM) produced by larger scale processes is a regulatory requirement. There is no cure for rabies but there is a vaccine to prevent it. FDA. Smith, T. R. F. et al. Optimally the same vaccine (either HDCV or PCEC) should be used throughout the vaccination series. While VLPs constitute self-associated protein units of well-defined sizes, the possibility of non-specific protein aggregation should be monitored by well-described hydrodynamic methods, e.g., light scattering based techniques. Formulation of two candidate sa-mRNA vaccines against Zika virus with a cationic nanoemulsion (CNE) has been reported to induce potent immunity in mice and non-human primates53. NIBSC. MethodsX 7. https://doi.org/10.1016/j.mex.2020.100806 (2020). J. Pharm. While ELISA, in various forms such as direct binding, competitive, or sandwich, has been a widely used and reliable technology, newer immunoassay technologies provide fast turn-around, high throughput, and good precision. Modified mRNA/lipid nanoparticle-based vaccines expressing respiratory syncytial virus F protein variants are immunogenic and protective in rodent models of RSV infection. Procedia Vaccinol. Measurements of A280nm should include corrections for light scattering that may result from protein aggregation. Maizel, J. V., White, D. O. FDA. Upset stomach. Commun. Tan, C. W. et al. Robust and faster turn-around assays for potency and other selected CQA will also be important for monitoring long-term and accelerated stability. Stabilization of human papillomavirus virus-like particles by non-ionic surfactants. 11, 3523 (2020). Reuters. Therefore, encapsulation efficiency should also be determined as a CQA. It is transmitted by a bite from a rabid animal (meaning an animal infected with rabies virus). Ther. The authors declare no competing interests. Lancet 397, 99111 (2021). Physicochemical and structural integrity of protein antigens are other CQA that can affect both potency and safety. A. C., van den Elzen, C., Ovelgnne, H., Baas, C. & Jongen, P. M. J. M. Estimation of the number of infectious measles viruses in live virus vaccines using quantitative real-time PCR. Dog Bite-Anti-Rabies Vaccine: . Recombinant viral vectored vaccines for human use. https://www.fda.gov/media/133757/download (2019). https://www.ema.europa.eu/en/human-regulatory/overview/public-health-threats/coronavirus-disease-covid-19/guidance-medicine-developers-other-stakeholders-covid-19 (2020). All available vaccine technology platforms, novel and traditional, are being utilized by different developers to produce vaccines against SARS-CoV-2. Google Scholar. Intracellular dsRNA produced during the replication cycle is a marker of RNA amplification. This is a part of Good Manufacturing Practice (GMP) and provides a safeguard against potentially costly and time-consuming clinical bridging. There are several examples of recombinant protein-based subunit and VLP antigens for which in vitro potency assays have been successfully developed. For a trimeric post-fusion F-protein-based Respiratory Syncytial Virus (RSV) vaccine, which was in clinical development (but not an approved product), the correlation between a sandwich ELISA IVRP and in vivo immunogenicity was established31, and IVRP was used as a lot release potency assay. Article J. Med. Only the rabies vaccines approved by the national health authorities should be used for the intra-dermal route. Two of the modified mRNA-based vaccines against SARS-CoV-2 have advanced rapidly through clinical phases58,59,60,61, and have recently delivered products that have received Emergency Use Authorization (EUA) by several regulatory agencies. In addition, clinical trials demonstrating acceptable safety profiles have been reported for all of the above-mentioned vectors62,63,64,65,66. Product-specific tests include in vitro bioanalytical batch release and characterization assays for the antigen and LNP. G.S. This assay is based on the indirect antibody labeling approach where protein expression proficiency measured by transfection of antigen encoded mRNA into Baby Hamster Kidney (BHK) cells followed by staining with fluorochrome tagged primary antibody52. However, safety concerns include reaching the balance between eliciting protective immune response and causing vaccine-induced infection. https://www.ema.europa.eu/en/medicines/human/EPAR/ervebo#authorisation-details-section (2019). EDQM. The European pharmacopoeia expert group (Group 15) has provided guidance on appropriate analytical strategies for viral vectored vaccines71 and, in addition, OCABR guidelines have been published for analyses of non-replicating human and chimpanzee adenovirus vectored vaccines21,22. mRNA antigens, on the other hand, are delivered directly into the cell cytoplasm for translation to the corresponding protein. PubMed Central https://www.nibsc.org/products/brm_product_catalogue/detail_page.aspx?catid=20/146 (2021). However, it is important to establish that focus formation is a result of infection and one is not observing simply a binding event that may not lead to CPE. However, if severe allergic reactions occur it may be advisable to switch to the alternate vaccine to complete the series. For drug product formulation and delivery, LNPs have been extensively used with the objective of stimulating innate immune response, and these studies include clinical trials54. A surrogate SARS-CoV-2 virus neutralization assay has been published, which is based on antibody-mediated inhibition of interaction between S-protein and angiotensin converting enzyme-2 (ACE-2) receptor35. Ranheim, T. et al. EMEA/H/C/005791/0000 (2021). Using QPCR to assign infectious potencies to adenovirus based vaccines and vectors for gene therapy: toward a universal method for the facile quantitation of virus and vector potency. In up to 99% of cases, domestic dogs are responsible for rabies virus transmission to humans. Immune responses elicited by different vaccines during the pre-clinical and clinical phases will be analyzed by implementing common protocols, antibody standards and equivalent key reagents across these laboratories92,93. . Vaccine 35, 19181925 (2017). Of the currently approved products for ebola, the recombinant VSV-based vaccine against the Zaire strain of ebola virus (rVSV-ZEBOV) utilizes a combination of infectivity determined by TCID50 assay and total viral particles assessed by droplet digital polymerase chain reaction (ddPCR) assay72. Reagents for in vitro immunochemical and biochemical assays, including the mAb CR3022, RBD of the S-protein, and human ACE2 are now being made available to eligible developers in collaborative arrangements with PATH and NIBSC. Development of a potent Zika virus vaccine using self-amplifying messenger RNA. Rabies vaccinations are highly effective, safe and well tolerated. https://doi.org/https://doi.org/10.1056/NEJMoa202 6920 (2020). J. Med. In addition, it is necessary, from a CMC standpoint, to monitor the concentration of replication competent viral particles that may appear during the manufacturing of a replication-deficient viral vector. CEPI. Rabies is a zoonotic viral disease which infects domestic and wild animals. Guideline for Pandemic COVID-19 Vaccine, Official Control Authority Batch Release of Pandemic COVID-19 Vaccine (Non-Replicating Human Adenovirus-Vectored Vaccine) (2020). Internet Explorer). Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. Two injections (each contains 0.1 ml of reconstituted vaccine) are given at two different sites per visit as intradermal injections on days 0, 3, 7 and 28. During development, leading up to a final process, detailed structural characterization provides a strong foundation for structure-function correlation for protein antigens. Exclusive: New global lab network will compare COVID-19 vaccines head-to-head (2020). Moderna COVID-19 Vaccine Emergency Use Authorization Review Memorandum. CAS Bloomberg. Even for one developer using a particular platform, multiple sites around the globe will be used to enable the manufacturing and rapid distribution of hundreds of millions of doses in different parts of the world. Sweeney, J. In addition, expression of the target antigen encoded by the inserted gene in an appropriate cell line is expected as a potency assay. provided global portfolio analysis, contributed immunology content, helped with drafting and editing, and provided literature information. For inactivated virus vaccines, the extent of inactivation is measured by following loss of infectious titer, while an immunoassay against a key epitope of the virus, which is still able to bind a specific mAb or polyclonal sera, can be used as a dose as well as a potency assay. The COVID-19 vaccine development landscape. While potency is intricately related to dose, quantitative methods to measure these attributes may be depending on the platform. There is a fundamental difference in the mechanism of action between pDNA and mRNA antigens. These are not all of the side effects that may occur. Thus, setting and meeting specifications for quality indicating assays are of paramount importance in ensuring delivery of safe and effective vaccines of consistent quality worldwide. Google Scholar. An mRNA vaccine against SARS-CoV-2preliminary report. These formulations typically involve encapsulation of mRNA in LNP. A rabies vaccine (human diploid cell) for human use, Freeze-dried produced by Chengdu Kanghua Biological Products Co.,Ltd is used to prevent human rabies. Article For DNA and RNA antigens, dose is readily measured by absorbance and fluorescence methods, or by quantitative PCR (qPCR). Jackson, L. A. et al. A SARS-CoV-2 surrogate virus neutralization test based on antibody-mediated blockage of ACE2spike proteinprotein interaction. Direct measurements of stability of secondary structures and thermal unfolding of protein antigens can be monitored by far-UV circular dichroism (CD) and differential scanning calorimetry (DSC). 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