US20210403879A1 - Method for inactivating zika virus and related methods - Google Patents
Method for inactivating zika virus and related methods Download PDFInfo
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- US20210403879A1 US20210403879A1 US16/768,550 US201816768550A US2021403879A1 US 20210403879 A1 US20210403879 A1 US 20210403879A1 US 201816768550 A US201816768550 A US 201816768550A US 2021403879 A1 US2021403879 A1 US 2021403879A1
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Definitions
- FIG. 19 shows the serum viremia of individual AG129 mice three days post-challenge, reported as PFU/mL.
- FIG. 25 shows compiled kinetics of inactivation data. Data compares infectious potency (TCID50) to RNA copy, and completeness of inactivation (COI) for samples from the four toxicology lots. These data indicate that the sensitivity of the COI assay is greater than TCID50.
- WO 97/37000 and WO 97/37001 describe the production of animal cells and cell lines that are capable of growth in suspension and in serum free media and are useful in the production and replication of viruses.
- the cells used for growing the at least one virus are Vero cells.
- the viral inoculum and the viral culture are preferably free from (i.e. will have been tested for and given a negative result for contamination by) herpes simplex virus, respiratory syncytial virus, parainfluenza virus 3, SARS coronavirus, adenovirus, rhinovirus, reoviruses, polyomaviruses, birnaviruses, circoviruses, and/or parvoviruses (WO 2006/027698).
- the numerical result of the multiplication of the formaldehyde concentration as measured in % (w/v) with the period of incubation with formaldehyde as measured in days is 0.025 to 0.5 and the formaldehyde concentration is 0.01% (w/v).
- the Zika virus preparation is treated with 0.008 to 0.015% (w/v) formalin for nine to eleven days at a temperature of 22° C. In some embodiments, the Zika virus preparation is treated with 0.008 to 0.015% (w/v) formalin for ten days at a temperature of 22° C. In some embodiments, the Zika virus preparation is treated with 0.01% (w/v) formalin for eight to twelve days at a temperature of 22° C. In some embodiments, the Zika virus preparation is treated with 0.01% (w/v) formalin for nine to eleven days at a temperature of 22° C. In some embodiments, the Zika virus preparation is treated with 0.01% (w/v) formalin for ten days at a temperature of 22° C.
- the vaccines and/or immunogenic compositions of the present disclosure containing one or more antigens from at least one inactivated Zika virus may be useful for treating or preventing Zika virus infection in a subject in need thereof and/or inducing an immune response, such as a protective immune response, against Zika virus in a subject in need thereof.
- the mixture of 50 parts of the composition comprising a virus which has been treated with formaldehyde with 1 part of 20% phosphoric acid and 2.5 parts of 1.0 mg/ml DNPH is incubated at a temperature of 18° C. to 30° C. for 10 to 30 minutes. In some embodiments the mixture of 50 parts of the composition comprising a virus which has been treated with formaldehyde with 1 part of 20% phosphoric acid and 2.5 parts of 1.0 mg/ml DNPH is incubated at a temperature of 18° C. to 30° C. for 15 to 25 minutes.
- the step of buffer exchange and/or dilution of the sample comprising the Zika virus preparation involves cross-flow filtration.
- cross-flow filtration which is also called tangential flow filtration the feed flow travels tangentially across the surface of the filter and not into the filter.
- the potency was determined by two different methods, TCID50 and plaque titration.
- the TCID50 was calculated by visual inspection of CPE (microscope) and by measuring the difference in absorbance (A 560 -A 420 ) of the wells displaying CPE (yellow in color) compared with red (no CPE). The plates were read on a plate reader, and applied to the same calculator as the microscopically read-plates (absorbance). The values in TCID50 between the two scoring techniques are quite similar, while the values obtained by plaque titration are lower.
- COI assay control The titer and back titration controls for this assay were performed using Vero indicator cells and scored in a TCID 50 96-well format with wells scored positive based on the media color change from pink to yellow, as a surrogate for cell death, or the presence of CPE.
- Virus titer control test Two independent replicates of the control virus (PRVABC59) of known titer were subjected to a 10-fold dilution series in media containing 2% FBS, and 100 ⁇ L of each dilution was added to four wells of a 96-well plate containing Vero cells.
- the method was validated in terms of specificity, linearity, accuracy, repeatability, intermediate precision, robustness, and stability of the sample.
- accuracy study the Zika vaccine drug product and aluminum hydroxide gel solution were spiked with a specific amount of formaldehyde, and the sample was mixed well by vortex before following the procedure described in Section 2.3.
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US11478541B2 (en) | 2017-11-03 | 2022-10-25 | Takeda Vaccines, Inc. | Method for inactivating Zika virus and for determining the completeness of inactivation |
US11975062B2 (en) | 2017-11-30 | 2024-05-07 | Takeda Vaccines, Inc. | Zika vaccines and immunogenic compositions, and methods of using the same |
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KR102253190B1 (ko) * | 2018-07-18 | 2021-05-18 | (주)진매트릭스 | 지카 바이러스 변이주와 이를 포함한 지카 백신 조성물 |
KR20220007077A (ko) * | 2019-05-08 | 2022-01-18 | 다케다 백신즈 인코포레이티드 | 불활성화 바이러스 조성물 및 지카 백신 제형 |
RU2717993C1 (ru) * | 2019-10-22 | 2020-03-27 | Федеральное бюджетное учреждение науки "Государственный научный центр вирусологии и биотехнологии "Вектор" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека (ФБУН ГНЦ ВБ "Вектор" Роспотребнадзора) | Способ получения антигена вируса Зика, обладающего иммуногенными и антигенными свойствами |
WO2023154043A1 (en) | 2022-02-09 | 2023-08-17 | Takeda Vaccines, Inc. | Zika vaccines and immunogenic compositions, and methods of using the same |
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