基于减毒和灭活病毒的疫苗
病毒疫苗包括灭活的病毒疫苗和减毒活病毒疫苗。这两种疫苗均依赖于传统的方法生产，即该致病病毒已经被发现并在生物反应器中进行了生产及纯化。这种类型的策略已经成功用于预防多种疾病，目前大多数预防病毒性疾病的疫苗均使用该策略生产。然而，该方法也有它需要克服的挑战：
- 确保病毒能够在生物反应器中生产
- 寻找到正确的宿主细胞
- 设计对每种疾病定制工艺
- 符合高生物安全水平下的生产要求，以保护操作员和环境
我们，赛多利斯，理解疫苗研发者以及病毒疫苗生产者的需求，并已经开发了专门的解决方案工具箱。
灭活病毒疫苗
灭活病毒疫苗在疫苗市场中占有重要的地位，其中以流感和脊髓灰质炎疫苗为主。除了新疫苗开发，工艺转移、产能扩大和连续改善也能在这一领域带来积极的改变，采用技术开发更安全、更高效的工艺，有助于降低总体成本。
我们邀请您发现赛多利斯专用于灭活病毒疫苗生产和纯化的技术工具箱。在您探索包膜病毒（例如SARS-CoV-2）、非包膜病毒和特定病毒（例如流感和脊髓灰质炎疫苗）的通用工艺概览时，您可点击每个步骤查看制造商在灭活病毒疫苗工作中面临的挑战——并发现赛多利斯如何提供独特的解决方案来满足这些需求。您还将看到有关工艺步骤可用技术的相关文献。
通用灭活病毒
细胞系/菌株选择
细胞系/菌株选择
Selecting cell lines and strains that produce commercially acceptable yields can be difficult and labor-intensive. Proper high throughput development tools — or a capable partner who has access to those tools — allow operators to confidently select a stable and safe cell line.
▶ Suspension Process
Need | Solution |
|---|---|
Selecting the optimum cell line or clone for the target virus relies on low-volume screening tools and fast analytics Fully scalable HTPD tools, inline analytics are needed to quickly define the best operating conditions for cell expansion (media selection, process parameters optimization) and virus propagation (media, feed, multiplicity of infection, time of infection, harvest time, etc.) Many in industry are moving away from serum-containing media in favor of chemically defined (CD) media to support process safety and batch consistency, Ultimately, having an experienced partner capable of providing the characterization tools for the cell line is the way to select a stable and safe cell line with confidence | The flexibility and functionality of Ambr® 15 & Ambr® 250 can accelerate cell line selection and process optimization. Ambr® is the only high-throughput cell culture system with scalable, multiparallel mini-bioreactors and automated functionality.
Additional Solutions to Consider:
|
▶ Adherent Process
Need | Solution |
|---|---|
Selecting the optimum cell line or clone for the target virus relies on low-volume screening tools and fast analytics Fully scalable HTPD tools, inline analytics are needed to quickly define the best operating conditions for cell expansion (media selection, microcarrier selection, process parameters optimization) and virus propagation (media, feed, multiplicity of infection, time of infection, harvest time, etc.) In order to effectively evaluate adaptation of adherent cell line to suspension, there is a pressing need for low volume screening tools, fast analytics Many in industry are moving away from serum-containing media in favor of chemically defined (CD) media to support process safety and batch consistency Ultimately, having an experienced partner capable of providing the characterization tools for the cell line is the way to select a stable and safe cell line with confidence | A new generation of Ambr® 15 tailored to microcarriers applications and Ambr® 250 vessels designed for microcarriers can accelerate cell line selection and process optimization. Ambr® is the only high-throughput cell culture system with scalable, multiparallel mini-bioreactors, automated functionality and vessels developed for microcarriers.
Additional Solutions to Consider:
|
Application Data:
种子扩增
种子扩增
Seed expansion from cell bank to production bioreactor must be successfully achieved before the virus propagation in the production bioreactor. The latest innovations use process intensification to reduce the seed train and ensure the highest possible process consistency while Process Analytical Technologies enable full process control.
▶ Suspension Process
Need | Solution |
|---|---|
Adjusting the seed expansion step to production scale relies on fully scalable technologies from process development to manufacturing Intensifying processes are critical to reduce the seed train and decrease cost of production while achieving higher cell concentration The sterility of the seed train relies on a closed process from the shake flask through the rocking motion bioreactor Process Analytical Technologies can ensure the robustness and control of the seed train | Biostat® RM, a fully GMP compliant, wave-mixed bioreactor, is an easy-to-use system for seed expansion offering the widest range of PAT in a Biosafety Level 2 environment
Additional Solutions to Consider:
|
Application Data:
- Single-Use, Stirred-Tank Bioreactors: Efficient Tools for Process Development and Characterization
▶ Adherent Process
Need | Solution |
|---|---|
Adjusting the seed expansion step to production scale relies on fully scalable technologies from process development to manufacturing Evaluation and implementation of the correct parameters and mixing conditions is critical to avoid cell detachment from the microcarriers and microcarriers breakage Process Analytical Technologies can ensure the robustness and control of the seed train | Biostat® RM, a fully GMP compliant, wave-mixed bioreactor, is an easy-to-use system for seed expansion offering the widest range of PAT in a Biosafety Level 2 environment
Additional Solutions to Consider:
|
病毒增殖
病毒增殖
Robust virus propagation in vaccine development relies on scalability from process development to production scale manufacturing while ensuring containment in a Biosafety Level 2/3 environment. The use of Process Analytical Technologies in viral-based vaccine applications enables process controls and monitoring to achieve a higher virus concentration.
▶ Suspension Process
Need | Solution |
|---|---|
Fully scalable technologies, from process development to manufacturing, are needed to take virus propagation to production scale Achieving higher virus concentration while decreasing cost of production relies on process intensification It is imperative to ensure containment and reduce contamination risk while working with infectious viral particles classified as Biosafety Level 2 Process Analytical Technologies in this environment are essential to enable robust processes and infection monitoring and control | Biostat STR® is a single use bioreactor offering the widest range of PAT in a Biosafety Level 2 environment, making it the perfect match for viral production:
Additional Solutions to Consider:
|
▶ Adherent Process
Need | Solution |
|---|---|
Fully scalable technologies, from process development to manufacturing, are needed to take virus propagation to production scale Evaluation and implementation of the correct parameters and mixing conditions is critical to avoid cell detachment from the microcarriers and microcarriers breakage It is imperative to ensure containment and reduce contamination risk while working with infectious viral particles classified as Biosafety Level 2 Process Analytical Technologies in this environment are essential to enable robust processes and infection monitoring and control | The Biostat STR® is a single use bioreactor, offering the widest range of PAT in a Biosafety Level 2 environment, making it the perfect match for viral production:
Additional Solutions to Consider:
|
Application data:
- Large pilot scale cultivation process study of adherent MDBK cells for porcine Influenza A virus propagation using a novel disposable stirred-tank bioreactor
- Comparative study of bluetongue virus serotype 8 production on BHK-21 cells in a 50L Biostat® STR single-use bioreactors vs roller bottles
- Inactivated Enterovirus 71 Vaccine Produced by 200-L Scale Serum-Free Microcarrier Bioreactor System Provides Cross-Protective Efficacy in Human SCARB2 Transgenic Mouse.
- Process intensification for Peste des Petites Ruminants Virus vaccine production
- Control and scale-up of a microcarrier-based viral vaccine process using BioPAT ® ViaMass for inline viable cell density measurement
澄清
澄清
Clarification of vaccines is a critical step that has a significant impact on product recovery and subsequent downstream purification. The main challenge of virus clarification is to recover a high yield of viral particles while efficiently removing cell debris, large aggregates and insoluble contaminants.
Clarification of Enveloped Viruses
▶ Suspension Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral particles are infectious, so containment during processing and disassembling is required to be compliant with Biosafety Level 2/3 requirements Enveloped viruses that are larger than 0.2µm make sterile filtration impossible, so the only way to ensure process sterility and operator safety in those circumstances is through pre-sterilized and closed solutions A high capacity clarification solution is needed to address the overall challenges of working with high cell density in a suspension process | Ksep® single use centrifuge is the leading single-use centrifuge for clarification from low to high cell density.
Additional Solutions to Consider:
|
▶ Adherent Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral particles are infectious, so containment during processing and disassembling is required to be compliant with Biosafety Level 2/3 requirements Enveloped viruses that are larger than 0.2µm make sterile filtration impossible, so the only way to ensure process sterility in those circumstances is through pre-sterilized and closed solutions A separation solution dedicated to microcarriers is required in order to remove cells growing on them | Sartopure® PP3 are high capacity pre-filters dedicated to non-intensified processes, providing the highest virus recovery
Additional Solutions to Consider:
|
Clarification of Non-Enveloped Viruses
▶ Suspension Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viruses are live particles, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2/3 requirements The significant number of contaminants released during cell lysis including DNA and host cell proteins (HCPs) leads to filter blockage, which can only be mitigated by a high capacity clarification solution A high capacity clarification solution also is needed to address the overall challenges of working with high cell density in a suspension process | Clarification is a challenging unit operation for suspension and lytic viral processes. A single step is often not sufficient and multiple technologies must be tested and combined, including flocculation, filter trains, and single-use centrifugation A wide range of technical approaches can be optimized and sequenced to meet broad product requirements Sartorius offers a range of filters to meet different needs:
|
Application Data:
Poster: Clarification of Adenovirus serotype 5: Robust protection of downstream purification steps
Poster: Implementation of a Process-Scale Adenovirus Purification with a Single-Use Platform
▶ Adherent Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral particles are infectious, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2/3 requirements The significant number of contaminants released during cell lysis including DNA and host cell proteins (HCPs) leads to filter blockage, which can only be mitigated by a high capacity clarification solution A separation solution dedicated to microcarriers is required in order to remove any cells growing on them | Sartopure® PP3 are high capacity pre-filters dedicated to non-intensified processes, providing the highest virus recovery
Additional Solutions to Consider:
|
Application data:
- Comparative study of bluetongue virus serotype 8 production on BHK-21 cells in a 50L Biostat® STR single-use bioreactors vs roller bottles
生物负载减低
生物负载减低
Bioburden reduction relies on membrane filtration of the clarified viral vaccine in order to protect the subsequent Downstream Unit Operations. A large range of membrane types and potential combinations create options to achieve the highest throughput while maintaining the highest recovery.
Need | Solution |
|---|---|
Viruses are large particles, and while most of them can be filtered through sterilizing grade filters (0.2µm), some cannot – if the virus is not filterable, it is important to evaluate 0.2µm (sterile filtration) as well as 0,45µm (bioburden reduction) and select fully closed and pre-sterilized technologies for the entire downstream process. Viral particles tend to adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Viral particles are infectious and must be processed in an environment compliant to Biosafety Level 2/3, so containment is critical | Maxicaps® MR, a ready-to-use, pre-sterilized, pre-assembled and self-contained filtration device allows connection of up to nine capsules in parallel for large-scale filtration operations
Additional Solutions to Consider:
|
Application Data:
Clarification of Adenovirus serotype 5: Robust protection of downstream purification steps
浓缩/透析
浓缩/透析
Concentration of viruses, contaminant removal and buffer exchange steps are essential steps in all viral vaccine processing, but these steps can be especially challenging when working with shear-sensitive viruses. The latest innovations combine low-shear operations and fully closed processing to allow the highest recovery with full containment – creating an ideal environment for viral vaccine manufacturing.
Need | Solution |
|---|---|
Viruses are large particles that are easy to separate from small contaminants. The optimization of the operating conditions should consider the benefit of contaminant removal (host cell proteins, DNA) which requires large molecular weight cut-off membrane cassettes and high throughput process development systems Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Prior to inactivation, viral particles are pathogenic, so containment is critical The significant number of contaminants including DNA and host cell proteins in addition to virus can foul filter membranes prompting the need for low-adsorption membranes Enveloped viral particles are fragile and may be damaged by shear exposure, so low shear systems and optimal cassette designs need to be used. | Hydrosart®, a state-of-the-art crossflow membrane designed for low adsorption of viruses has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral-based vaccines
Additional Solutions to Consider:
|
病毒捕获层析
病毒捕获层析
Virus capture is a critical step in the Downstream process, removing the majority of contaminants while concentrating the virus. Innovative chromatography membrane formats eliminate barriers to scalability and allow implementation of high capacity membranes at very large scale.
Need | Solution |
|---|---|
Viruses are large particles that cannot diffuse into chromatography beads, and can be trapped in small pores leading to low recovery during elution, requiring alternatives to traditional bead-based chromatography media Addressing an array of viruses requires a wide range of chromatography media and operating conditions with screening to ensure both highest capacity and highest recovery of viruses, so high throughput screening tools and a variety of chromatography ligands are needed Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Prior to inactivation, viral particles are pathogenic, so containment is critical | Sartobind® 8mm chromatography membranes offer the widest range of ligands and a unique cassette scale-up solution up to 100L of membrane volume, ideal for virus purification in bind and elute mode
Additional Solutions to Consider:
|
Application data:
Optimizing Adenovirus Purification Processes Using Sartobind® Q and STIC PA anion exchange membranes
Poster: Implementation of a Process-Scale Adenovirus Purification with a Single-Use Platform
Membrane Chromatography Cassettes for Bind & Elute Applications of Viruses and Large Proteins
精纯层析
精纯层析
A second step of chromatography for polishing might be required to remove remaining contaminants, such as DNA and HCP. Membrane adsorbers allow the easy implementation of this secondary chromatography step in a fully contained format.
Need | Solution |
|---|---|
Viral processes can lead to cell lysis, releasing a significant number of contaminants including DNA that has to be removed in downstream processes without impact on the overall virus recovery – something that can be achieved through high recovery chromatographic solutions for polishing in flowthrough mode Viruses are large particles that can be trapped in small pores, leading to low recovery during flowthrough operations, requiring alternatives to traditional bead-based chromatography media Addressing an array of viruses requires a wide range of chromatography media and operating conditions with screening to ensure both maximum levels of impurity removal and highest recovery of viruses, so high throughput screening tools and diverse chromatography ligands are needed. Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & pre-sterilized systems Prior to inactivation, viral particles are pathogenic, so containment is critical | Sartobind® 4mm chromatography membranes offer the widest range of ligands and a unique cassette scale-up solution up to 100L of membrane volume, ideal for contaminant removal in flowthrough mode
Additional Solutions to Consider:
|
Additional Data:
Optimizing Adenovirus Purification Processes Using Sartobind® Q and STIC PA anion exchange membranes
Poster: Implementation of a Process-Scale Adenovirus Purification with a Single-Use Platform
Rational Development of Two Flowthrough Purification Strategies for Adenovirus Type 5 and Retro Virus-Like Particles
灭活/裂解
灭活/裂解
Inactivation of the virus is critical to ensure safety for patients. Inactivation is achieved by exposing viruses to chemical agents, such as formalin and beta-propiolactone. State-of-the-art single use mixing solutions offer a robust inactivation, while ensuring containment for operator safety.
Need | Solution |
|---|---|
In order to ensure the inactivation of every viral particle, having drop-free mixing, no dead volumes and no adsorption of chemicals is essential Containment in this process is critical for two reasons: viral particles are pathogenic prior to inactivation and toxic chemicals are used during the inactivation process. It is important to verify the compatibility of films with the structure and degradation profiles of the chemicals being used Viral particles are fragile and may be damaged by shear exposure, so low shear mixing solutions are needed. | The Flexsafe® Pro Mixer technology, the only single-use mixing solution with integrated sensors, accommodates both low-shear and rapid mixing applications in a fully closed design which enables viral inactivation while preserving the virus
Additional Solutions to Consider:
|
浓缩/透析
浓缩/透析
Concentration of viruses, contaminant removal and buffer exchange steps are essential steps in all viral vaccine processing, but these steps can be especially challenging when working with shear-sensitive viruses. The latest innovations combine low-shear operations and fully closed processing to allow the highest recovery with full containment – creating an ideal environment for viral vaccine manufacturing.
Need | Solution |
|---|---|
Viruses are large particles that are easy to separate from small contaminants. The optimization of the operating conditions should consider the benefit of contaminant removal (host cell proteins, DNA) which requires large molecular weight cut-off membrane cassettes and high throughput process development systems Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Prior to inactivation, viral particles are pathogenic, so containment is critical The significant number of contaminants including DNA and host cell proteins in addition to virus can foul filter membranes prompting the need for low-adsorption membranes Enveloped viral particles are fragile and may be damaged by shear exposure, so low shear systems and optimal cassette designs need to be used. | Hydrosart®, a state-of-the-art crossflow membrane designed for low adsorption of viruses has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral-based vaccines
Additional Solutions to Consider:
|
除菌过滤
除菌过滤
The final sterile filtration step is critical to prevent microbial contamination and ensure patient safety. Innovative risk mitigation strategies ensure safety of the complete solution, from sterile filtration to storage in single-use bags.
Need | Solution |
|---|---|
Viruses are large particles, and while most of them can be filtered through sterilizing grade filters (0.2µm), some cannot. It is critical to evaluate 0.2µm (sterile filtration) as well as 0,45µm (bioburden reduction) and select fully closed and pre-sterilized technologies across the entire downstream process if a virus is not filterable Viral particles adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Viral particles are infectious so containment is critical | Helium supplier integrity test and point-of-use leak test with Sartocheck® is the best risk mitigation strategy across the single-use bag portfolio, offering assurance of integrity when containment and product sterility are critical
Additional Solutions to Consider:
|
其他技术
病毒定量，即在规定体积内测量病毒颗粒浓度，是开发病毒载体时所需要的宝贵信息，要求尽可能地实现实时测量。现有的技术均有局限性，因此需要采用互补性的新技术来确保快速且准确的定量。
Need | Solution |
|---|---|
Virus infectivity tests can take days or weeks to get results – rapid detection analytics are needed to address that critical issue ELISA and PCR methods are faster than infectivity tests, but there is a higher incidence of error variability since they measure virus building blocks and derive total titers from these numbers, leading to a need for alternative, reliable analytics Situations often demand ready-to-use analytics, but ELISA and PCR need development and validation time to ensure method robustness A lot of virus samples are generated during process development and manufacturing of viral based vaccines, highlighting the need for an easier workflow allowing rapid quantification of virus samples | The Virus Counter® Platform, a unique and rapid virus quantification method, is the only system able to directly quantify assembled viruses (protein and nucleic acids) for real-time insight
|
先进的化学计量学方法，例如实验设计（DOE）和多变量数据分析（MVDA），最大限度可视化数据，不仅能节约成本外，同时还能提高工艺的可靠性和稳健性。
需求 | 解决方案 |
|---|---|
质量源于设计（QbD）依赖于DoE来了解关键工艺参数，而后者取决于很多实验的实施——这个工具可帮助设计和规划实验，以帮助减轻负担 工艺分析技术产生了大量难以分析的数据，因此对评价历史数据的工具的需求巨大，以便识别相关性、进行故障排除并获得工艺理解 大多数疫苗开发商和制造商并不是统计学家，而是依赖于用户友好型界面和软件 | Umetrics®套件包括三种为工艺开发人员和制造商设计的用户友好型直观软件解决方案，以支持独特的数据可视化、广泛的向导功能和可定制绘图期间的数据分析，从而最大限度地提高可用性和多功能性
These software solutions are fully integrated in most Sartorius systems and are available as stand-alone programs |
采用日益发展且集成一次性传感器的一次性技术，不仅能实时收集和分析数据，还能提高工艺效率、降低污染风险、提高操作人员的安全性并提高总体产品产量。
| 需求 | 解决方案 |
|---|---|
Vaccine manufacturers want to better understand and control their processes, which are very complex and highly regulated, but to do so requires real-time measurement of critical process parameters to allow process monitoring, set-point control (feed control and bleed control), event time point predictions (harvest and infection) and timely identification of process deviations A range of PAT sensors would be required to effectively apply QbD principles and ensure consistency in product quality and quantity, quickly identifying and correcting process deviations to reduce the risk of lost batches The ability to integrate PAT into single-use systems would mitigate risk of spillages and contamination during sampling | BioPAT® toolbox, an expending range of fully qualified single-use sensors, are integrated across the Sartorius portfolio making Sartorius the market leader for PAT
|
Application Data:
减毒活病毒疫苗
由于分子生物学和基因工程领域的快速发展，新一代的减毒活病毒疫苗也即将问世。针对流感等病毒的疫苗，以及登革热或呼吸道合胞病毒（RSV）等较新的应用，都在这一前景广阔的研究领域范围内。快速发展领域通常需要创新技术来跟上新的工艺，以不断提高安全性和生产率。
我们邀请您发现赛多利斯专用于灭活病毒疫苗生产和纯化的技术工具箱。在您探索包膜病毒（例如SARS-CoV-2）、非包膜病毒和特定病毒（例如流感和脊髓灰质炎疫苗）的通用工艺概览时，您可点击每个步骤查看制造商在灭活病毒疫苗工作中面临的挑战——并发现赛多利斯如何提供独特的解决方案来满足这些需求。您还将看到有关工艺步骤可用技术的相关文献。
通用减毒病毒
细胞系/菌株选择
细胞系/菌株选择
Selecting cell lines and strains that produce commercially acceptable yields can be difficult and labor-intensive. Proper high throughput development tools — or a capable partner who has access to those tools — allow operators to confidently select a stable and safe cell line.
▶ Suspension Process
Need | Solution |
|---|---|
Selecting the optimum cell line or clone for the target virus relies on low-volume screening tools and fast analytics Fully scalable HTPD tools, inline analytics are needed to quickly define the best operating conditions for cell expansion (media selection, process parameters optimization) and virus propagation (media, feed, multiplicity of infection, time of infection, harvest time, etc.) Many in industry are moving away from serum-containing media in favor of chemically defined (CD) media to support process safety and batch consistency, Ultimately, having an experienced partner capable of providing the characterization tools for the cell line is the way to select a stable and safe cell line with confidence | The flexibility and functionality of Ambr® 15 & Ambr® 250 can accelerate cell line selection and process optimization. Ambr® is the only high-throughput cell culture system with scalable, multiparallel mini-bioreactors and automated functionality.
Additional Solutions to Consider:
|
▶ Adherent Process
Need | Solution |
|---|---|
Selecting the optimum cell line or clone for the target virus relies on low-volume screening tools and fast analytics Fully scalable HTPD tools, inline analytics are needed to quickly define the best operating conditions for cell expansion (media selection, microcarrier selection, process parameters optimization) and virus propagation (media, feed, multiplicity of infection, time of infection, harvest time, etc.) In order to effectively evaluate adaptation of adherent cell line to suspension, there is a pressing need for low volume screening tools, fast analytics Many in industry are moving away from serum-containing media in favor of chemically defined (CD) media to support process safety and batch consistency Ultimately, having an experienced partner capable of providing the characterization tools for the cell line is the way to select a stable and safe cell line with confidence | A new generation of Ambr® 15 tailored to microcarriers applications and Ambr® 250 vessels designed for microcarriers can accelerate cell line selection and process optimization. Ambr® is the only high-throughput cell culture system with scalable, multiparallel mini-bioreactors, automated functionality and vessels developed for microcarriers.
Additional Solutions to Consider:
|
Application Data:
- Ambr® 15 microcarrier
- Ambr® 250 microcarrier
- Ambr® 250 microcarrier poster
种子扩增
种子扩增
Seed expansion from cell bank to production bioreactor must be successfully achieved before the virus propagation in the production bioreactor. The latest innovations use process intensification to reduce the seed train and ensure the highest possible process consistency while Process Analytical Technologies enable full process control
▶ Suspension Process
Need | Solution |
|---|---|
Adjusting the seed expansion step to production scale relies on fully scalable technologies from process development to manufacturing Intensifying processes are critical to reduce the seed train and decrease cost of production while achieving higher cell concentration The sterility of the seed train relies on a closed process from the shake flask through the rocking motion bioreactor | Biostat® RM, a fully GMP compliant, wave-mixed bioreactor, is an easy-to-use system for seed expansion offering the widest range of PAT in a Biosafety Level 2 environment
Additional Solutions to Consider:
|
Application Data:
Garcia L, Mourino M, Urniza A, Juanola S. BioProcess J, 2019; 18.
Evaluation of a Single-Use Benchtop Process Development System to Optimize Cell Growth and Scale-Up of Veterinary Vaccine Production
▶ Adherent Process
Need | Solution |
|---|---|
Adjusting the seed expansion step to production scale relies on fully scalable technologies from process development to manufacturing Evaluation and implementation of the correct parameters and mixing conditions is critical to avoid cell detachment from the microcarriers and microcarriers breakage The sterility of the seed train relies on a closed process from the shake flask through the rocking motion bioreactor Process Analytical Technologies can ensure the robustness and control of the seed train | Biostat® RM,, a fully GMP compliant, wave-mixed bioreactor, is an easy-to-use system for seed expansion offering the widest range of PAT in a Biosafety Level 2 environment
Additional Solutions to Consider:
|
Application Data:
- Single-Use, Stirred-Tank Bioreactors: Efficient Tools for Process Development and Characterization
病毒增殖
病毒增殖
Robust virus propagation in vaccine development relies on scalability from process development to production scale manufacturing while ensuring containment in a Biosafety Level 2/3 environment. The use of Process Analytical Technologies in viral-based vaccine applications enables process controls and monitoring to achieve a higher virus concentration
▶ Suspension Process
Need | Solution |
|---|---|
Fully scalable technologies, from process development to manufacturing, are needed to take virus propagation to production scale Achieving higher virus concentration while decreasing cost of production relies on process intensification It is imperative to ensure containment and reduce contamination risk while working with infectious viral particles classified as Biosafety Level 2 Process Analytical Technologies in this environment are essential to enable robust processes and infection monitoring and control | Biostat STR® is a single use bioreactor offering the widest range of PAT in a Biosafety Level 2 environment, making it the perfect match for viral production:
Additional Solutions to Consider:
|
▶ Adherent Process
Need | Solution |
|---|---|
Fully scalable technologies, from process development to manufacturing, are needed to take virus propagation to production scale It is imperative to ensure containment and reduce contamination risk while working with infectious viral particles classified as Biosafety Level 2 Evaluation and implementation of the correct parameters and mixing conditions is critical to avoid cell detachment from the microcarriers and microcarriers breakage Process Analytical Technologies in this environment are essential to enable robust processes and infection monitoring and control | The Biostat STR® is a single use bioreactor, offering the widest range of PAT in a Biosafety Level 2 environment, making it the perfect match for viral production:
Additional Solutions to Consider:
|
Application data:
- Large pilot scale cultivation process study of adherent MDBK cells for porcine Influenza- A virus propagation using a novel disposable stirred-tank bioreactor
- Comparative study of bluetongue virus serotype 8 production on BHK-21 cells in a 50L Biostat® STR single-use bioreactors vs roller bottles
- Inactivated Enterovirus 71 Vaccine Produced by 200-L Scale Serum-Free Microcarrier Bioreactor System Provides Cross-Protective Efficacy in Human SCARB2 Transgenic Mouse.
- Process intensification for Peste des Petites Ruminants Virus vaccine production
- Control and scale-up of a microcarrier-based viral vaccine process using BioPAT ® ViaMass for inline viable cell density measurement
澄清
澄清
Clarification of vaccines is a critical step that has a significant impact on product recovery and subsequent downstream purification. The main challenge of virus clarification is to recover a high yield of viral particles while efficiently removing cell debris, large aggregates and insoluble contaminants
Clarification of Enveloped Viruses
▶ Suspension Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Attenuated viruses are live particles, so containment during processing and disassembling is required to be compliant with Biosafety Level 2/3 requirements Enveloped viruses that are larger than 0.2µm make sterile filtration impossible, so the only way to ensure process sterility in those circumstances is through pre-sterilized and closed solutions High capacity clarification solution is needed to address the overall challenges of working with high cell density in a suspension process Enveloped viral particles are fragile may be damaged by shear exposure during clarification, so low shear solutions are essential to protect the infectious nature of the virus | Ksep® single use centrifuge is the leading single-use centrifuge for clarification from low to high cell density.
Additional Solutions to Consider:
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▶ Adherent Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Attenuated viruses are live particles, so containment during processing and disassembling is required to be compliant with Biosafety Level 2/3 requirements Enveloped viruses that are larger than 0.2µm make sterile filtration impossible, so the only way to ensure process sterility in those circumstances is through pre-sterilized and closed solutions A separation solution dedicated to adherent processes is required in order to remove microcarriers Enveloped viral particles are fragile may be damaged by shear exposure during clarification, so low shear solutions are essential to protect the infectious nature of the virus | Sartopure® PP3 are high capacity pre-filters dedicated to non-intensified processes, providing the highest virus recovery.
Additional Solutions to Consider:
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Clarification of Non-Enveloped Viruses
▶ Suspension Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Attenuated viruses are live particles, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2/3 requirements The significant number of contaminants released during cell lysis including DNA and host cell proteins (HCPs) leads to filter blockage, which can only be mitigated by a high capacity clarification solution High capacity clarification solution also is needed to address the overall challenges of working with high cell density in a suspension process | Clarification is a challenging unit operation for suspension and lytic viral processes. A single step is often not sufficient and multiple technologies must be tested and combined, including flocculation, filter trains, and single-use centrifugation A wide range of technical approaches can be optimized and sequenced to meet broad product requirements Sartorius offers a range of filters to meet different needs:
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▶ Adherent Process
Need | Solution |
|---|---|
Viral particles adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Attenuated viruses are live particles, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2/3 requirements The significant number of contaminants released during cell lysis including DNA and host cell proteins (HCPs) leads to filter blockage, which can only be mitigated by a high capacity clarification solution A separation solution dedicated to adherent processes is required in order to remove microcarriers | Sartopure® PP3 are high capacity pre-filters dedicated to non-intensified processes, providing the highest virus recovery
Additional Solutions to Consider:
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Application data:
- Comparative study of bluetongue virus serotype 8 production on BHK-21 cells in a 50L Biostat® STR single-use bioreactors vs roller bottles
生物负载减低
生物负载减低
Bioburden reduction relies on membrane filtration of the clarified viral vaccine in order to protect the subsequent Downstream Unit Operations. A large range of membrane types and potential combinations create options to achieve the highest throughput while maintaining the highest recovery.
Need | Solution |
|---|---|
Viruses are large particles, and while most of them can be filtered through sterilizing grade filters (0.2µm), some cannot – if the virus is not filterable, it is important to 0,45µm (bioburden reduction) and select fully closed and pre-sterilized technologies for the entire downstream process. Viral particles tend to adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Attenuated viruses are live particles and must be processed in an environment compliant to Biosafety Level 2, so containment is critical | Maxicaps® MR, a ready-to-use, pre-sterilized, pre-assembled and self-contained filtration device allows connection of up to nine capsules in parallel for large-scale filtration operations
Additional Solutions to Consider:
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浓缩/透析
浓缩/透析
Concentration of viruses, contaminant removal and buffer exchange steps are essential steps in all viral vaccine processing, but these steps can be especially challenging when working with shear-sensitive viruses. The latest innovations combine low-shear operations and fully closed processing to allow the highest recovery with full containment – creating an ideal environment for viral vaccine manufacturing
Need | Solution |
|---|---|
Viruses are large particles that are easy to separate from small contaminants. The optimization of the operating conditions should consider the benefit of contaminant removal (host cell proteins, DNA) which requires large molecular weight cut-off membrane cassettes and high throughput process development systems Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Attenuated viruses are live particles that must be processed in an environment compliant to Biosafety Level 2, so containment is critical The significant number of contaminants including DNA and host cell proteins in addition to virus can foul filter membranes prompting the need for low-adsorption membranes | Hydrosart®, a state-of-the-art crossflow membrane designed for low adsorption of viruses has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral-based vaccines
Additional Solutions to Consider:
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病毒捕获层析
病毒捕获层析
Virus capture is a critical step in the Downstream process, removing the majority of contaminants while concentrating the virus. Innovative chromatography membrane formats eliminate barriers to scalability and allow implementation of high capacity membranes at very large scale
Need | Solution |
|---|---|
Viruses are large particles that cannot diffuse into chromatography beads, and can be trapped in small pores leading to low recovery during elution, requiring alternatives to traditional bead-based chromatography media Diversity of viruses requires a wide range of chromatography media and operating conditions with screening to ensure both highest capacity and highest recovery of infectious particle, so high throughput screening tools and a variety of chromatography ligands are needed Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Attenuated viruses are live particles that must be processed in an environment compliant to Biosafety Level 2, so containment is critical | Sartobind® 8mm chromatography membranes offer the widest range of ligands and a unique cassette scale-up solution up to 100L of membrane volume, ideal for virus purification in bind-and-elute mode
Additional Solutions to Consider:
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Application data:
精纯层析
精纯层析
A second step of chromatography for polishing might be required to remove remaining contaminants, such as DNA and HCP. Membrane adsorbers allow the easy implementation of this secondary chromatography step in a fully contained format
Need | Solution |
|---|---|
Viral processes can lead to cell lysis, releasing a significant number of contaminants including DNA that has to be removed in downstream processes without impact on the overall virus recovery – something that can be achieved through high recovery chromatographic solutions for polishing in flowthrough mode Viruses are large particles that cannot diffuse into chromatography beads, and can be trapped in small pores leading to low recovery during elution, requiring alternatives to traditional bead-based chromatography media Diversity of viruses requires a wide range of chromatography media and operating conditions with screening to ensure both highest capacity and highest recovery of infectious particle, so high throughput screening tools and diverse chromatography ligands are needed. Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Attenuated viruses are live particles that must be processed in an environment compliant to Biosafety Level 2, so containment is critical | Sartobind® 4mm chromatography membranes offer the widest range of ligands and a unique cassette scale-up solution up to 100L of membrane volume, ideal for contaminant removal in flowthrough mode
Additional Solutions to Consider:
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浓缩/透析
浓缩/透析
Concentration of viruses, contaminant removal and buffer exchange steps are essential steps in all viral vaccine processing, but these steps can be especially challenging when working with shear-sensitive viruses. The latest innovations combine low-shear operations and fully closed processing to allow the highest recovery with full containment – creating an ideal environment for viral vaccine manufacturing
Need | Solution |
|---|---|
Viruses are large particles that are easy to separate from small contaminants. The optimization of the operating conditions should consider the benefit of contaminant removal (host cell proteins, DNA) which requires large molecular weight cut-off membrane cassettes and high throughput process development systems Some viruses are larger than 0.2µm and cannot be sterile filtered, so those circumstances require closed & sterile environments Attenuated viruses are live particles that must be processed in an environment compliant to Biosafety Level 2, so containment is critical The significant number of contaminants including DNA and host cell proteins in addition to virus can foul filter membranes prompting the need for low-adsorption membranes | Hydrosart®, a state-of-the-art crossflow membrane designed for low adsorption of viruses has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral-based vaccines
Additional Solutions to Consider:
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除菌过滤
除菌过滤
The final sterile filtration step is critical to prevent microbial contamination and ensure patient safety. Innovative risk mitigation strategies ensure safety of the complete solution, from sterile filtration to storage in single-use bags
Need | Solution |
|---|---|
Viruses are large particles, and while most of them can be filtered through sterilizing grade filters (0.2µm), some cannot. It is critical to evaluate 0.2µm (sterile filtration) as well as 0,45µm (bioburden reduction) and select fully closed and pre-sterilized technologies across the entire downstream process if a virus is not filterable Viral particles adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Attenuated viruses are live particles that must be processes in an environment compliant to Biosafety Level 2, so containment is critical | Helium supplier integrity test and point-of-use leak test with Sartocheck® is the best risk mitigation strategy across the single-use bag portfolio, offering assurance of integrity when containment and product sterility are critical
Additional Solutions to Consider:
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其他技术
病毒定量，即在规定体积内测量病毒颗粒浓度，是开发病毒载体时所需要的宝贵信息，要求尽可能地实现实时测量。现有的技术均有局限性，因此需要采用互补性的新技术来确保快速且准确的定量。
Need | Solution |
|---|---|
Virus infectivity tests can take days or weeks to get results – rapid detection analytics are needed to address that critical issue ELISA and PCR methods are faster than infectivity tests, but there is a higher incidence of error variability since they measure virus building blocks and derive total titers from these numbers, leading to a need for alternative, reliable analytics Situations often demand ready-to-use analytics, but ELISA and PCR need development and validation time to ensure method robustness A lot of virus samples are generated during process development and manufacturing of viral based vaccines, highlighting the need for an easier workflow allowing rapid quantification of virus samples | The Virus Counter® Platform, a unique and rapid virus quantification method, is the only system able to directly quantify assembled viruses (protein and nucleic acids) for real-time insight
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先进的化学计量学方法，例如实验设计（DOE）和多变量数据分析（MVDA），最大限度可视化数据，不仅能节约成本外，同时还能提高工艺的可靠性和稳健性。
需求 | 解决方案 |
|---|---|
质量源于设计（QbD）依赖于DoE来了解关键工艺参数，而后者取决于很多实验的实施——这个工具可帮助设计和规划实验，以帮助减轻负担 工艺分析技术产生了大量难以分析的数据，因此对评价历史数据的工具的需求巨大，以便识别相关性、进行故障排除并获得工艺理解 大多数疫苗开发商和制造商并不是统计学家，而是依赖于用户友好型界面和软件 | Umetrics®套件包括三种为工艺开发人员和制造商设计的用户友好型直观软件解决方案，以支持独特的数据可视化、广泛的向导功能和可定制绘图期间的数据分析，从而最大限度地提高可用性和多功能性
These software solutions are fully integrated in most Sartorius systems and are available as stand-alone programs |
采用日益发展且集成一次性传感器的一次性技术，不仅能实时收集和分析数据，还能提高工艺效率、降低污染风险、提高操作人员的安全性并提高总体产品产量。
| 需求 | 解决方案 |
|---|---|
Vaccine manufacturers want to better understand and control their processes, which are very complex and highly regulated, but to do so requires real-time measurement of critical process parameters to allow process monitoring, set-point control (feed control and bleed control), event time point predictions (harvest and infection) and timely identification of process deviations A range of PAT sensors would be required to effectively apply QbD principles and ensure consistency in product quality and quantity, quickly identifying and correcting process deviations to reduce the risk of lost batches The ability to integrate PAT into single-use systems would mitigate risk of spillages and contamination during sampling | BioPAT® toolbox, an expending range of fully qualified single-use sensors, are integrated across the Sartorius portfolio making Sartorius the market leader for PAT
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Application Data: