Good manufacturing practice for sterile manufacturing
无菌生产技术的生产质量管理规范
Q1：Are antibiotics used in all vaccine manufacturing processes?
Q1：所有疫苗生产过程中都使用抗生素吗?
A1：Certain antibiotics may be used in vaccine production to help prevent bacterial contamination during manufacturing. As a result, small amounts of antibiotics may be present in some of the vaccines, either in very small amounts, or even in undetectable quantities.
Some antibiotics can cause severe allergic reactions in some children, such as hives, swelling at the back of the throat, and low blood pressure. However, antibiotics most likely to cause severe allergic reactions, e.g. penicillins, cephalosporins and sulfonamide drugs, are not used in vaccine production and, therefore, are not contained in vaccines. Examples of antibiotics used during vaccine manufacture include neomycin, polymyxin B, streptomycin and gentamicin.
A1：某些抗生素可用于疫苗生产，以帮助防止生产过程中的细菌污染。因此，一些疫苗中可能含有少量抗生素，有的含量非常少，有的甚至无法检测到。
一些抗生素会在一些儿童中引起严重的过敏反应，如荨麻疹、喉头水肿和低血压。然而，最可能引起严重过敏反应的抗生素，例如青霉素、头孢菌素和磺胺类药物，并未用于疫苗生产，因此也不包含在疫苗中。疫苗生产中使用的抗生素的例子包括新霉素、多粘菌素B、链霉素和庆大霉素。
Q2：Does vaccine manufacturing require a dedicated facility?
Q2：疫苗生产是否需要专用设施?
A2：There are requirements for specific vaccines (tetanus, BCG – since they are spore formers) to have dedicated facilities, but not for others. WHO Technical Report Series 986, Annex 2, and WHO Technical Report Series 999, Annex 2, mention the possibility of manufacturing multiple products in the same facilities.
However, considerations should include the specificity of the installations, the long processing times, and the extremely high validation requirements when thinking about multi-purpose facilities, especially when there is an alternance between live and inactivated vaccines.
A2：对特定疫苗(破伤风疫苗、卡介苗——因为它们形成芽孢)需要专门设施，但对其他疫苗没有要求。世卫组织技术报告系列986附件2和世卫组织技术报告系列999附件2，提到了在同一设施中生产多种产品的可能性。
然而，在考虑多功能设施时，特别是在活疫苗和灭活疫苗之间存在交替时，应考虑到设施的特殊性、长处理时间和极高的验证要求。
Q3：In addition to temperature and time, is pressure also a part of the sterilization condition?
Q3：除了温度和时间，压力也是灭菌条件的一部分吗?
A3：There is a correlation between temperature and pressure, for example, in an autoclave. However, pressure is equally distributed, but the temperature may not be.
A3：温度和压力之间是有关联的，例如，在高压灭菌器中，压力是均匀分布的，但温度可能不是。
Q4：What should be an acceptable bioburden level prior to the sterile filtration?
Q4：产品灭菌过滤前可接受的生物负荷水平是多少?
A4：Prior to final sterilizing filtration, products must show a bioburden lower than 10 CFU/100 ml.
A4：在产品最终灭菌过滤之前，生物负荷必须低于10 CFU/100毫升。
Q5：What is the meaning of integrity testing of sterilizing filters?
Q5：灭菌过滤器完整性检测的意义是什么?
A5：Integrity testing means to test if the filter is intact, normally before and certainly after filtration.
A5：完整性测试是指测试过滤器是否完整，通常在过滤前和过滤后。
Q6：When is the right time to test the integrity of the filter before use or before or after sterilization?
Q6：什么时候检验过滤器的完整性是正确的，在使用前还是灭菌前或灭菌后?
A6：For final filtration, the integrity testing should be done after sterilization as filter properties may have been impacted by sterilization. However, there may be some flexibility for early-stage sterile filtration.
A6：最后过滤时，应在灭菌后进行完整性测试，因为灭菌可能会影响过滤器的性能。然而，早期灭菌过滤可能有一定的灵活性。
Q7：Should pre-use post-sterilization integrity testing (PUPSIT) be tested before use after a certain holding time or tested then used after 5 days, for example?
Q7：使用前灭菌后完整性测试(PUPSIT)应在使用前进行测试，还是在使用一定时间，例如5天后进行测试?
A7：It is acceptable to test after a certain holding time (e.g., 5 days), if you can validate that the filter is still sterile after 5 days. However, it is recommended to sterilize, cool down and test immediately.
A7：在使用一定时间(例如5天)后进行测试是可以接受的，如果您可以验证过滤器在5天后仍然是无菌的。但是，建议立即灭菌、冷却和测试。
Q8：What is the recommendation for wetting agent for PUPSIT?
Q8：对使用前灭菌后完整性测试的润湿剂有什么建议?
A8：When the integrity test is done before the filtration, it is recommended to use the product itself.
A8：当完整性测试在过滤前完成时，建议使用产品本身。
Q9：What is the function of flush bags?
Q9：冲洗袋的作用是什么?
A9：Flush bags are there to collect fluid used to wet the filter before the test.
A9：冲洗袋用于收集测试前用来湿润过滤器的液体。
Q10：How do we maintain sterility of the filter if we do integrity testing prior to use?
Q10：如果我们在使用前做了完整性测试，我们如何保持过滤器的无菌性?
A10：The integrity testing is done from the unsterile side (upstream) of the filter. The air used is sterile filtered, and the solution used to wet the filter also comes from the upstream side of the filter.
A10：完整性测试是从过滤器的未消毒侧(上游)进行的。所使用的空气经过无菌过滤，用于湿润过滤器的溶液也来自过滤器的上游侧。
Q11：Is hydrogen peroxide bio- decontamination process validation required?
Q11：是否需要过氧化氢生物净化(灭菌)工艺验证?
A11：Yes, there should be validation runs with bioindicators to test if micro-organisms have been effectively killed.
A11：是的，应该进行生物指标验证，以测试微生物是否已被有效杀死。
Q12：What are the room classification requirements for activities in the manufacturing of sterile products?
Q12：无菌产品生产活动的房间分类要求是什么?
A12：Room grades and allowed/prescribed operations are presented in GMP guidelines such as WHO good manufacturing practice (GMP) for sterile products, Annex 1 of EU GMP, and Annex 1 of PIC/S GMP.
A12：房间等级和允许/规定的操作在药品生产质量管理规范指南中有介绍，如WHO无菌产品药品生产质量管理规范、欧盟药品生产质量管理规范附件1和国际药品认证合作组织药品生产质量管理规范附件1。
Q13：How acceptable is it if the area classification is based on an International Organization for Standardization (ISO) standard only without consideration
for viable/microbial quality of the environment?
Q13：如果区域分级是基于国际标准化组织(ISO)的标准，而没有考虑环境的可生存/微生物质量，这是否可以接受?
A13：It is not acceptable. For pharmaceuticals, there is a need to consider viable and non- viable particles.
A13：这是不可接受的。对于药品来说，有必要考虑可存活的和不可存活的粒子。
Q14：Can the isolator for filling sterile products (class A) be installed in a class C room?
Q14：灌装A类无菌产品的隔离器能安装在C类房间吗?
A14：Yes, that is the advantage of isolators; class D is also possible, but considerations should also be on the number of ancillary isolator- compatible accessories.
A14：是的，这就是隔离器的优点；D类也可以，但还应该考虑辅助隔离器兼容附件的数量。
Q15：Is it possible to use restricted access barrier systems (RABSs) installed in Class D rooms to fill aseptic products?
Q15：是否可能使用安装在D级房间的限制进入屏障系统(RABSs)来灌装无菌产品?
A15：Aseptic filling takes place in Class A under Class B background. The Class D environment is only possible when there is an isolator.
A15：无菌灌装在A级和B级背景下进行。D级环境只有在有隔离器的情况下才可能出现。
Q16：Are non-viable and viable counts required during the transfer of working cell expansion in flasks when we do it under Laminar Air Flow (LAF)?
Q16：在层流（LAF）条件下，用烧瓶进行工作细胞增值和扩张转移时，是否需要非活性颗粒和活性颗粒计数？
A16：Both non-viable and viable counts are required if the process is done under Grade A.
A16：如果工艺是在A级以下进行的，则非活菌计数和活菌计数都是必需的。
Q17：What should we do when monitoring shows deviations in critical control parameters like particles and microbial loads in aseptic facilities?
Q17：当监测显示关键控制参数(如无菌设施中的颗粒和微生物负荷)出现偏差时，我们应该怎么做?
A17：Consider stopping operations, evaluate the criticality (excursions or massive deviations) and investigate the cause, and identify the germs.
A17：考虑停止操作，评估危险程度(偏移或巨大偏差)，调查原因，确定细菌。
Q18：What is the link between closed system and quality by design (QbD)?
Q18：封闭系统与设计质量(QbD)之间的联系是什么?
A18：Closed systems are certainly a part of QbD, as they reduce greatly the risk of contamination.
A18：封闭系统当然是系统和质量设计的一部分，因为它们大大降低了污染的风险。
Q19：What is the purpose of the smoke test?
Q19：烟雾测试的目的是什么?
A19：The smoke test can be used to visualize airflows (e.g. show laminarity).
A19：烟雾测试可用于可视化气流(例如显示层流性)。
Q20：Can sterilization of vials and stoppers be done together in one step?
Q20：西林瓶灭菌和瓶塞灭菌可以同时进行吗?
A20：It cannot be done in one step since procedures are essentially different: dry heat depyrogenation for vials; and steam sterilization for stoppers.
A20：这不能一步完成，因为程序本质上是不同的:西林瓶用干热灭菌；瓶塞用蒸汽灭菌。
Q21： What are the requirements in terms of monitoring/testing for the release of sterile gowns to be used in a controlled environment (grades A or B) when those are obtained from a supplier?
Q21：在受控环境(A级或B级)中，从供应商处获得的无菌防护服放行的监测/检测要求是什么?
A21：For sterile gowns from a supplier, the requirement should include audits of the supplier, and focus on packaging and sterilization procedures, as well as leak testing on packaging.
A21：对于供应商提供的无菌防护服，要求应包括对供应商的审核，重点是包装和灭菌程序，以及包装上的气密测试。
Q22：How to manage breakdowns or stoppages in the sterilization area where the production is running?
Q22：如何处理生产过程中发生的故障或停产?
A22：Stops and breakdowns should be simulated during media fill tests. To prevent stoppages, preventive maintenance, an uninterruptible power supply, and good equipment should be considered. However, when breakdowns or stoppages occur, the following actions include: recording it in the batch manufacturing records; considering media fill simulation; removing and discarding part of the batch left open during the stoppage time, etc.
A22：在培养基灌装试验期间，应模拟停机和故障。为了防止停工，应考虑定期维修、不间断的电源供应和良好的设备。但是，当发生故障或停工时，应采取以下措施：将故障记在此批生产记录中；考虑培养基灌装模拟实验；对在停工时间内批次中未封闭的部分进行报废处理等。
Q23：Which water type do we use for vaccine production – water for injection (WFI) or purified water (PW)?
Q23：我们生产疫苗使用哪一种水——注射用水(WFI)还是纯净水(PW)?
A23：Water type depends on the process step, but at the final stage, it will be WFI.
A23：水的类型取决于工序，但在最后阶段，将是注射用水。
Q24：Does a manufacturer have to wait for inactivated test results for the product to go to the next step in the inactivated area, or can they be based on historical data?
Q24：制造商是否需要等待灭活测试结果才能进入下一个灭活区域，或者他们可以基于历史数据来进行?
A24：If the process is properly validated, the process can continue, taking into account that there is a risk; it depends on how long it takes to verify the inactivation.
A24：如果生产进程通过适当的验证，考虑到存在的风险，加工程序可以继续；这取决于验证灭活的时间。
Q25：Does biosafety level (BSL) decrease after inactivation?
Q26：灭活后生物安全等级(BSL)是否降低?
A25：Yes, because the product does not contain any bio-active agent anymore.
A25：是的，因为该产品不再含有任何生物活性剂。
Q26：Should media fill be based on the product with the largest batch size?
Q26：培养基灌装是否应以最大批量的产品为准?
A26：It is not so much related to the batch size, but the number of filled units. If the batch size is smaller than 5000 units, a complete batch is filled. Otherwise, the minimum number is 5000, although many companies opt for larger numbers. The minimum numbers are defined in GMP guidelines such as Annex 1 of the EU GMP, PIC/S GMP, WHO guidelines, and the United States guideline on aseptic processing.
A26：这与批量大小没有太大关系，而是与灌装单位的数量有关。如果批量小于5000个单位，则会灌装一个完整的批次。此外，最低数量是5000，尽管许多公司选择更大的数量。最低数量在药品生产质量管理规范的指南中有定义，如欧盟药品生产质量管理规范附件1、国际药品认证合作组织药品生产质量管理规范、WHO指南和美国无菌工艺指南。
Q27：Is it necessary to do media fills for the last step of bulk manufacturing such as sterile filtration of bulk?
Q27：是否有必要在原液生产的最后一步(如原液无菌过滤)进行培养基灌装?
A27：The media fill should reflect all steps of the bulk where sterility is at risk. For example, filtration of sterile media.
A27：培养基灌装会影响原液的所有步骤，其中无菌性存在风险。例如无菌培养基的过滤。
Q28：If there is a deviation that causes the holding time between formulation and filling to be longer than the usual process, should it be simulated on the next period of media fill?
Q28：如果有偏差导致配方和灌装之间的持续时间比常规程序要长，是否应该在下一阶段的培养基灌装中进行模拟?
A28：When holding times are validated, it is important to plan a worst-case scenario to avoid deviations at a later stage. However, if the situation exists, the
longer holding time would be considered at the next media fill as a validation exercise.
A28：当确认了持续时长后，重要的是要计划出最坏的情况，以避免在后期出现偏差。然而，如果存在这种情况，较长的持续时间将被认为是下一个培养基灌装的验证练习。
Q29：Is a fumigation schedule necessary in vaccine manufacturing?
Q29：疫苗生产是否需要进行熏蒸?
A29：Generally, there is no need for scheduled fumigation. Good householding practices should be sufficient.
A29：一般不需要设定熏蒸。良好的生产管理规范就足够了。
Q30：When a single-use system (SUS) is used for fill and finish products, do we have to do the assembling under laminar flow? and is one filter before filling (close to the point of fill) enough?
Q30：当一次性系统(SUS)用于灌装和成品时，我们必须在层流下进行组装吗？在灌装(接近灌装点)之前用一个过滤器是否足够?
A30：The use of a laminar flow for connections depends on whether there are conventional aseptic connections or connectors. Sterile filtration near the point of fill is necessary. However, the filling itself (including assembly of the filling elements) is prone to contamination as well; thus, the filtration is not a guarantee of sterility.
A30：使用层流连接取决于是常规无菌连接还是连接器。在灌装点附近无菌过滤是必要的。然而，灌装本身(包括灌装元件的装配)也容易受到污染; 因此，过滤并不能保证无菌。
Q31：Is it necessary to do aseptic process validation for seed preparation?
Q31：是否有必要对种子制备进行无菌工艺验证?
A31：Yes, because the purity of the seed is critical; a contaminated seed will negatively affect all subsequent process steps.
A31：是的，因为种子的纯度很关键；被污染的种子会对后续的所有加工步骤不利。
Q32： Is there any guideline specifying whether we need isolator systems for product filling, or is a RABS acceptable? or is it based on quality risk management (QRM)?
Q32：是否有相关指南说明产品灌装是否需要隔离系统，或者可否采用限制进入屏障系统？还是基于质量风险管理(QRM)?
A32：The decision is both QRM-based and costbased, as isolators are expensive as are their ancillary features, e.g., Realtime Transport Protocol (RTP) ports, transfer systems, etc.
A32：这个决定是以质量风险管理和成本为基础，因为隔离系统和其他辅助功能一样昂贵，如实时传输协议(RTP)端口、传输系统等。
Q33：For replication-incompetent vectors of COVID-19 vaccine, what are the suggested containment strategies for the whole production process starting from cell culture up to formulation and filling?
Q33：对于复制不完全载体的COVID-19疫苗，从细胞培养到配方和灌装的整个生产过程中建议采取哪些控制策略?
A33：According to the current requirement, the manufacturer has to make their own safety risk assessment (see WHO Laboratory Biosafety Manual, 4th edition).
A33：根据目前的要求，制造商必须进行自己的安全风险评估(见WHO实验室生物安全手册第四版)。
Q34：What are the most critical steps for inspection for fill and finish vaccine manufacturers?
Q34：对疫苗灌装和成品的制造商来说，进行检查的最关键步骤是什么?
A34: Of particular importance is quality assurance (QA), and the manufacture of sterile products must strictly follow carefully established and validated methods of manufacture and control. A contamination control strategy (CCS) should be implemented across the facility in order to define all critical control points and assess the effectiveness of all the controls (design, procedural, technical and organizational) and monitoring measures employed to manage risks associated with contamination.
The CCS should be actively updated and should drive continuous improvement of manufacturing and control methods.
A34:尤其重要的是质量保证(QA)，无菌产品的生产必须严格遵循详细规定和验证的生产控制方法。污染控制策略(CCS)应在整个设施中实施，以确定所有关键控制点，并评估所有控制(设计、程序、技术和组织)的有效性，以及用于管理污染相关风险的监测措施。
污染控制策略应及时更新，并应推动生产和控制策略的持续改进。