病毒載體疫苗
病毒載體疫苗為生物制藥行業(yè)帶來新的希望，解決了很多未滿足的適應(yīng)癥。病毒載體領(lǐng)域采用40多種載體研究基因治療和疫苗，其中腺病毒和改良的安卡拉痘苗病毒最常用于疫苗應(yīng)用，腺相關(guān)病毒通常用于基因治療，而慢病毒通常用于基因改造的細(xì)胞療法。為了充分發(fā)揮病毒載體疫苗的潛力，很多生產(chǎn)商正在尋求創(chuàng)新的技術(shù)，以為更多產(chǎn)、強(qiáng)化和靈活的工藝提供支持。
病毒載體疫苗平臺
了解病毒載體為什么是疫苗生產(chǎn)中最常用的平臺之一。
病毒載體
我們邀請您了解賽多利斯專用于病毒載體生產(chǎn)和純化的技術(shù)解決方案。在您探索包膜載體、非包膜載體和常用載體（例如腺病毒、AAV、慢病毒和MVA）的通用工藝概覽時(shí)，您可點(diǎn)擊每個(gè)步驟查看生產(chǎn)商在病毒載體疫苗開發(fā)工作中面臨的挑戰(zhàn)——并發(fā)現(xiàn)賽多利斯如何提供獨(dú)特的解決方案來滿足這些需求。您還將看到有關(guān)工藝步驟可用技術(shù)的相關(guān)文獻(xiàn)。?
通用病毒載體疫苗
細(xì)胞系/菌株選擇
細(xì)胞系/菌株選擇
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 robustness 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:
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?? 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 with inline analytics are needed to quickly define the best operating conditions for cell expansion (media selection, microcarrier selection, process parameters optimization) and viral vector propagation by infection or plasmid transfection (media, feed, multiplicity of infection, time of infection, plasmid concentration, inducibility 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 and 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:
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種子擴(kuò)增
種子擴(kuò)增
Seed expansion from cell bank to production bioreactor must be successfully achieved before the viral vector 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:
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? 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 microcarrier 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:
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病毒增殖
病毒增殖
Robust viral vector propagation in vaccine development relies on scalability from process development to production scale manufacturing while ensuring containment in a Biosafety Level 2 environment. The use of Process Analytical Technologies in viral vector applications enables process controls and monitoring to achieve a higher viral vector 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:
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? Adherent Process
Need | Solution |
|---|---|
Fully scalable technologies, from process development to manufacturing, are needed to take viral vector propagation to production scale Evaluation and implementation of the correct parameters and mixing conditions is critical to avoid cell detachment from the microcarriers and? microcarrier breakage It is imperative to ensure containment and reduce contamination risk while working with viral vectors, which are live viruses 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:
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澄清
澄清
Clarification of viral vectors is a critical step that has a significant impact on product recovery and subsequent downstream purification. The main challenge of viral vector clarification is to recover a high yield of viral vector particles while efficiently removing cell debris, large aggregates and insoluble contaminants.
Clarification of Enveloped Viruses
? Suspension Process?
Need | Solution |
|---|---|
Viral vectors adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral vectors are live particles, so containment during processing and disassembling is required to be compliant with Biosafety Level 2/3 requirements Enveloped viral vectors 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 Enveloped viral vectors 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 vectors adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral vectors are live particles, so containment during processing and disassembling is required to be compliant with Biosafety Level 2 requirements Enveloped viral vectors 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 Enveloped viral vectors are fragile and may be damaged by shear exposure during clarification so low shear solutions are essential to ensure they remain infectiou | Sartopure? PP3 are high capacity pre-filters dedicated to non-intensified processes, providing the highest recovery of viral particles in combination with lowest filtration costs among polypropylene pre-filters available on the market
Additional Solutions to Consider: ?
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Clarification of Non-Enveloped Viruses
? Suspension Process
Need | Solution |
|---|---|
Viral vectors adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral vectors are live particles, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2 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:
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? Adherent Process
Need | Solution |
|---|---|
Viral vectors adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Viral vectors are live particles, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2 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 recovery of viral particles combined with the lowest filtration costs among polypropylene pre-filters available on the market
Additional Solutions to Consider:
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生物負(fù)載減低
生物負(fù)載減低
Bioburden reduction relies on membrane filtration of the clarified viral vector 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 |
|---|---|
Viral vectors 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 vectors tend to adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Viral vectors 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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? Application Data:
濃縮/透析
濃縮/透析
Concentration of viral vectors, contaminant removal and buffer exchange steps are essential steps in all viral vector-based vaccine processing, but these steps can be especially challenging when working with shear-sensitive viral vectors. 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 vector manufacturing.
Need | Solution |
|---|---|
Viral vectors 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 viral vectors are larger than 0.2μm and cannot be sterile filtered, so those circumstances require closed & sterile environments Viral vectors are live viruses that must be compliant to Biosafety Level 2, so containment is critical The significant number of contaminants including DNA and host cell proteins in addition to viral vectors adsorb to membranes, prompting the need for low-adsorption membranes Enveloped viral vectors 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 viral vectors has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral-vector based vaccines
?Additional Solutions to Consider:
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Application Data:
病毒捕獲層析
病毒捕獲層析
Viral vector capture is a critical step in the Downstream process, removing the majority of contaminants while concentrating the viral vectors. Innovative chromatography membrane formats eliminate barriers to scalability and allow implementation of high capacity membranes at very large scale.
Need | Solution |
|---|---|
Viral vectors 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 viral vectors requires a wide range of chromatography media and operating conditions with screening to ensure both highest capacity and highest recovery of infectious particles, so high throughput screening tools and a variety of chromatography ligands are needed Some viral vectors are larger than 0.2μm and cannot be sterile filtered, so those circumstances require closed & pre-sterilized systems Viral vectors are live particles and 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 viral vector purification in bind and elute mode
Additional Solutions to Consider:
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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
精純層析
精純層析
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 vector 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 Viral vectors 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 viral vectors requires a wide range of chromatography media and operating conditions with screening to ensure both maximum levels of impurity removal and highest recovery of infectious particles, so high throughput screening tools and diverse chromatography ligands are needed. Some viral vectors are larger than 0.2μm and cannot be sterile filtered, so those circumstances require closed & pre-sterilized systems Viral vectors are live particles and 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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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
- Rational Development of Two Flowthrough Purification Strategies for Adenovirus Type 5 and Retro Virus-Like Particles
濃縮/透析
濃縮/透析
Concentration of viral vectors, contaminant removal and buffer exchange steps are essential steps in all viral vector-based vaccine processing, but these steps can be especially challenging when working with shear-sensitive viral vectors. 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 vector manufacturing.
Need | Solution |
|---|---|
Viral vectors 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 viral vectors are larger than 0.2μm and cannot be sterile filtered, so those circumstances require closed & sterile environments Viral vectors are live viruses that must be compliant to Biosafety Level 2, so containment is critical The significant number of contaminants including DNA and host cell proteins in addition to viral vectors adsorb to membranes, prompting the need for low-adsorption membranes Enveloped viral vectors 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 viral vectors has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral-vector based vaccines
?Additional Solutions to Consider:
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Application Data:
除菌過濾
除菌過濾
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 |
|---|---|
Viral vectors 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 viral vector is not filterable Viral vectors adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Viral vectors are live particles and must be processed 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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其他技術(shù)
病毒載體定量，即在規(guī)定體積內(nèi)測量病毒載體顆粒濃度，是開發(fā)病毒載體時(shí)所需要的寶貴信息，需要盡可能接近實(shí)時(shí)測量值?，F(xiàn)有的技術(shù)均有局限性，因此需要采用互補(bǔ)性的新技術(shù)來確?？焖偾覝?zhǔn)確的定量。
| 需求 | 解決方案 |
|---|---|
病毒傳染性測試需要幾天或幾周的時(shí)間才能獲得結(jié)果，因此，需要快速的檢測分析來解決這一重要挑戰(zhàn) ELISA和PCR法比傳染性測試更快，但由于它們測量病毒的組成部分并從這些數(shù)字中得出總滴度，誤差變異性的發(fā)生率更高，因此需要替代的、可靠的分析方法 一般情況下需要現(xiàn)成的分析結(jié)果，但ELISA和PCR的開發(fā)和確認(rèn)需要花費(fèi)時(shí)間，才能確保方法的穩(wěn)健性 病毒載體工藝開發(fā)和制造期間會(huì)生成大量的病毒樣本，需要有更簡單的工作流程，才能快速定量病毒樣本 | Virus Counter?平臺是一種獨(dú)特、快速的病毒定量方法，它也是唯一一個(gè)能夠直接量化已組裝的病毒（蛋白質(zhì)和核酸），實(shí)時(shí)進(jìn)行分析的系統(tǒng)
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應(yīng)用數(shù)據(jù)：
- 在Virus Counter? 3100上使用基于抗體檢測的快速、實(shí)時(shí)定量慢病毒顆粒
- 具有高致病性潛力的病毒家族的快速總顆粒定量
先進(jìn)的化學(xué)計(jì)量學(xué)方法，例如實(shí)驗(yàn)設(shè)計(jì)（DOE）和多變量數(shù)據(jù)分析（MVDA）為數(shù)據(jù)提供了廣泛的可視性，除了節(jié)約成本外，最終還能提高工藝可靠性和穩(wěn)健性
| 需求 | 解決方案 |
|---|---|
質(zhì)量源于設(shè)計(jì)（QbD）依賴于DoE來了解關(guān)鍵工藝參數(shù)，而后者取決于很多實(shí)驗(yàn)的實(shí)施——這個(gè)工具可幫助設(shè)計(jì)和計(jì)劃實(shí)驗(yàn)，以幫助減輕負(fù)擔(dān) 工藝分析技術(shù)產(chǎn)生了大量難以分析的數(shù)據(jù)，因此對評價(jià)歷史數(shù)據(jù)的工具的需求巨大，以便識別相關(guān)性、進(jìn)行故障排除并獲得工藝?yán)斫?/p> 大多數(shù)病毒載體開發(fā)和生產(chǎn)人員并不是統(tǒng)計(jì)學(xué)家，而是依賴于用戶友好型界面和軟件 | Umetrics?套件包括三個(gè)專為工藝開發(fā)和生產(chǎn)人員設(shè)計(jì)的軟件，其易于使用且界面直觀，可通過獨(dú)特的數(shù)據(jù)可視化、大量的向?qū)K和可定制繪圖功能為數(shù)據(jù)分析提供支持，從而最大限度地提高可用性和通用性
這些軟件解決方案在大多數(shù)賽多利斯系統(tǒng)中完全集成，并可用作單機(jī)程序 |
Application Data:
隨著一次性技術(shù)的不斷發(fā)展，集成了一次性傳感器的一次性技術(shù)不僅實(shí)現(xiàn)了實(shí)時(shí)數(shù)據(jù)收集和分析，還提高了過程效率，降低了污染風(fēng)險(xiǎn)，提高了操作人員的安全性，并提高了整體滴度
需求 | 解決方案 |
|---|---|
病毒載體制造商想要更好地理解并控制其工藝，這些工藝非常復(fù)雜而且受到高度監(jiān)管，但是這樣做需要實(shí)時(shí)測量關(guān)鍵工藝參數(shù)，以允許工藝監(jiān)測、設(shè)定值控制（進(jìn)料控制和出料控制）、事件時(shí)間點(diǎn)預(yù)測（收獲和感染）以及工藝偏差的及時(shí)識別 需要一系列的PAT傳感器，以有效應(yīng)用QbD原則，并確保產(chǎn)品質(zhì)量和數(shù)量的一致性，快速識別和糾正工藝偏差，以降低批次丟失的風(fēng)險(xiǎn) 將PAT集成至一次性系統(tǒng)中的能力將降低采樣期間的泄露和污染風(fēng)險(xiǎn) | BioPAT?工具箱是一系列完全合格的一次性傳感器，其整合在賽多利斯產(chǎn)品組合中，使得賽多利斯成為PAT領(lǐng)域的市場領(lǐng)導(dǎo)者
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應(yīng)用數(shù)據(jù)：
- 使用BioPAT?ViaMass在線活細(xì)胞密度測量的微載體病毒疫苗工藝的控制和放大
- ?
基于腺病毒的疫苗
腺病毒是經(jīng)常用作病毒載體的非包膜DNA病毒，具備大量優(yōu)勢：
- ??? 易于進(jìn)行基因操控
- ??? 非復(fù)制性
- ??? 可接受較大轉(zhuǎn)基因DNA插入
- ??? 能產(chǎn)生穩(wěn)健的轉(zhuǎn)基因特異性T和B響應(yīng)
其最初被評價(jià)為基因治療載體，但現(xiàn)在大多應(yīng)用在疫苗以及溶瘤病毒。今天，有50%以上基于病毒載體的疫苗在使用腺病毒。市場上銷售的首個(gè)基于腺病毒的疫苗是埃博拉病毒疫苗。?
我們誠邀您了解賽多利斯專用于病毒載體生產(chǎn)和純化的技術(shù)解決方案。在您探索我們的通用腺病毒工藝概覽時(shí)，您可點(diǎn)擊每個(gè)步驟，查看制造商在腺病毒工作中面臨的挑戰(zhàn)——并了解賽多利斯如何提供獨(dú)特的解決方案來滿足這些需求。您還將看到有關(guān)工藝步驟的相關(guān)文獻(xiàn)。
腺病毒疫苗
細(xì)胞系/菌株選擇?
細(xì)胞系/菌株選擇?
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.
Need | Solution |
|---|---|
Selecting the optimum cell line or clone for the adenovirus relies on low-volume screening tools and fast analytics Fully scalable HTPD tools and inline analytics are needed to quickly define the best operating conditions for cell expansion (media selection, process parameters optimization) and adenovirus 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 in an effort 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 (usually HEK293) with confidenc | 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 and bioreactor system with scalable, multiparallel, automated functionality
Additional Solutions to Consider:
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種子擴(kuò)增
種子擴(kuò)增
Seed expansion from cell bank to production bioreactor must be successfully achieved before the viral vector 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 Technology enables full process control.
Need | Solution |
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Adjusting the seed propagation 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 ?
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病毒增殖
病毒增殖
Robust viral vector propagation in vaccine development relies on scalability from process development to production scale manufacturing while ensuring containment in a Biosafety Level 2 environment. The use of Process Analytical Technologies in viral vector applications enables process controls and monitoring to achieve a higher viral vector concentration.
Need | Solution |
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Fully scalable technologies, from process development to manufacturing, are needed to take virus propagation to production scale Achieving higher adenovirus 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, such as adenoviruses Process Analytical Technologies in this environment are essential to enable robust process 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 vector production:
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澄清
澄清
Clarification of viral vectors is a critical step that has a significant impact on product recovery and subsequent downstream purification. The main challenge of viral vector clarification is to recover a high yield of viral vector particles while efficiently removing cell debris, large aggregates and insoluble contaminants.
Need | Solution |
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Adenoviruses tend to adsorb to depth filters, so it is critical to remove cells or cell debris while maintaining high product recovery Adenoviruses are live virus particles, so containment during processing and disassembling is required to be in compliance with Biosafety Level 2 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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生物負(fù)載減低
生物負(fù)載減低
Bioburden reduction relies on membrane filtration of the clarified viral vector 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 |
|---|---|
Adenovirus particles tend to adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Adenoviruses are live particles 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
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Application Data:
濃縮/透析
濃縮/透析
Concentration of viral vectors, contaminant removal and buffer exchange steps are essential steps in all viral vector-based vaccine processing, but these steps can be especially challenging when working with shear-sensitive viral vectors. 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 vector manufacturing.
Need | Solution |
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Adenoviruses 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 Adenoviruses 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 adenovirus can foul filter membranes prompting the need for low-adsorption membranes | Hydrosart?, a state-of-the-art crossflow membrane designed for low adsorption of viral vectors has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral vector-based vaccines
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Application Data:
病毒捕獲層析
病毒捕獲層析
Viral vector capture is a critical step in the Downstream process, removing the majority of contaminants while concentrating the viral vectors. Innovative chromatography membrane formats eliminate barriers to scalability and allow implementation of high capacity membranes at very large scale.
Need | Solution |
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Adenoviruses 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 Purification of adenoviruses require operating conditions with screening to ensure both highest capacity and highest recovery of infectious particles on anion exchangers, so high throughput screening tools are needed Adenoviruses are live particles that must be processed in an environment compliant to Biosafety Level 2, so containment is critical | Sartobind? 8mm chromatography membranes, designed for adenovirus purification in bind and elute mode, offer the widest range of ligands and a unique cassette scale-up solution up to 100L of membrane volume
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精純層析
精純層析
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 |
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Adenovirus processes 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 adenovirus recovery – something that can be achieved through high recovery chromatographic solutions for polishing in flowthrough mode Adenoviruses are large particles can be trapped in small pores leading to low recovery during elution, requiring alternatives to traditional bead-based chromatography media Purification of adenoviruses requires a wide range of chromatography media and operating conditions with screening to ensure both highest impurity removal and highest recovery of infectious particles, so high throughput screening tools and diverse chromatography ligands are needed Adenoviruses 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
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Application Data:
- Optimizing Adenovirus Purification Processes Using Sartobind? Q and STIC PA anion exchange membranes
- 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
濃縮/透析
濃縮/透析
Concentration of viral vectors, contaminant removal and buffer exchange steps are essential steps in all viral vector-based vaccine processing, but these steps can be especially challenging when working with shear-sensitive viral vectors. 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 vector manufacturing.
Need | Solution |
|---|---|
Adenoviruses 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 Adenoviruses 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 adenovirus can foul filter membranes prompting the need for low-adsorption membranes | Hydrosart?, a state-of-the-art crossflow membrane designed for low adsorption of viral vectors has the benefit of resistance to chemicals, steam, heat and gamma-irradiation, allowing a wide range of operating conditions for viral vector-based vaccines
?Additional Solutions to Consider:
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Application Data:
除菌過濾
除菌過濾
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 |
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Adenovirus particles tend to adsorb to filters, so it is important to use non-adsorptive membranes as an alternative to maximize recovery Adenoviruses are live particles that must processed 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
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其他技術(shù)
先進(jìn)的化學(xué)計(jì)量學(xué)方法，例如實(shí)驗(yàn)設(shè)計(jì)（DOE）和多變量數(shù)據(jù)分析（MVDA），最大限度可視化數(shù)據(jù)，不僅能節(jié)約成本外，同時(shí)還能提高工藝的可靠性和穩(wěn)健性。
| 需求 | 解決方案 |
|---|---|
質(zhì)量源于設(shè)計(jì)（QbD）依賴于DoE來了解關(guān)鍵工藝參數(shù)，而后者取決于很多實(shí)驗(yàn)的實(shí)施——這個(gè)工具可幫助設(shè)計(jì)和計(jì)劃實(shí)驗(yàn)，以幫助減輕負(fù)擔(dān) PAT會(huì)生成大量難以分析的數(shù)據(jù)，因此，用于評價(jià)歷史數(shù)據(jù)的工具的需求大幅增長，旨在識別相關(guān)性、排除故障并對工藝有深入的理解 大多數(shù)疫苗開發(fā)和生產(chǎn)人員并不是統(tǒng)計(jì)學(xué)家，而是依賴于用戶友好型界面和軟件 | Umetrics?套件包括三個(gè)專為工藝開發(fā)和生產(chǎn)人員設(shè)計(jì)的軟件，其易于使用且界面直觀，可通過獨(dú)特的數(shù)據(jù)可視化、大量的向?qū)K和可定制繪圖功能為數(shù)據(jù)分析提供支持，從而最大限度地提高可用性和通用性。
這些軟件選項(xiàng)完全集成在大多數(shù)賽多利斯系統(tǒng)中，并可作為單機(jī)程序使用 |
隨著一次性技術(shù)的不斷發(fā)展，集成了一次性傳感器的一次性技術(shù)不僅實(shí)現(xiàn)了實(shí)時(shí)數(shù)據(jù)收集和分析，還提高了過程效率，降低了污染風(fēng)險(xiǎn)，提高了操作人員的安全性，并提高了整體滴度
| 需求 | 解決方案 |
|---|---|
Viral vector-based 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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