1、精選優質文檔-傾情為你奉上Process Validation (PV) Protocol XXXXXX工藝驗證方案Written by起 草 人Written date起草日期年 月 日Reviewed by審 核 人Reviewed date審核日期年 月 日Approved by批 準 人Approved date批準日期年 月 日Written department起草部門生效日期年 月 日Issued by:頒發部門： Copy number:拷 貝 號： Change record:Revision number變更記載： 修訂號 批準日期 生效日期 Change rea

2、son and goal: 變更原因及目的：Distribution department:分發部門:所長辦公室 人力資源部 后勤保衛部 物資供應部 工程服務部 生產管理部 質量保證部 銷 售 部 倉 儲 部 疫苗一室 疫苗二室 疫苗三室 疫苗五室 疫苗六室 生化制品室 細胞因子室 血清制品室 實驗動物室 分包裝室 培養基室 Verification department (department): 驗證科室（部門）： Verify completion date: 驗證完成日期： Validity: 有 效 期： Index 目錄1. Purpose目的This purpose o

3、f this process validation is to provide documented evidence to show that the combination of the personnel, materials, equipment, methods, environmental conditions and the other related utilities XXX Co., LTD (XXX) can ensure the consistent production of products conforming to the enterprise internal

4、 standards and the national standards, and the processes are reliable and conforming to the GMP requirements. The process validation and the test results are to be documented based on this validation protocol.本工藝驗證用于提供文件化的證據，證明XXX疫苗一室生產XXX原液的人員、材料、設備、方法、環境條件以及其它有關公用設施的組合可以始終如一的生產出符合企業內控標準及國家法定標準的產品，

5、工藝穩定可靠，符合GMP要求，工藝驗證的過程和檢查的結果將按照該驗證方案進行記錄。2. Scope范圍This protocol is applicable to the process validation of XXX produced by the equipment located XXX.本方案適用于XXX原液生產的工藝驗證。由于工作細胞庫制備的過程并非常規生產過程，因此，不在本次驗證中進行考察。The labeling and packaging processes are not within the scope of this document since these proc

6、esses have been validated per PQ protocol (labeling) and PQ protocol (packaging), and these processes have minimal impact on finished product quality. 貼標簽和包裝也不在此文件范圍內，因為這些工藝已經按照PQ方案（貼標簽）和PQ方案(包裝)驗證過，并且這些工藝對產品質量極少有影響。3. Responsibility職責3.1 AAA responsibilities AAA的職責 ü Compile Protocol方案的編寫ü

7、; Guide Protocol execution, data collection/ Provide test data for review upon request by customer指導方案的實施，數據的收集/在客戶要求時提供測試數據供其審核ü Final report Review/ Analyze and compile data into the summary report最終報告的審核/對數據進行分析并將其編輯入總結報告之中3.2 XXX responsibilities XXX的職責n Production department 生產部ü Revi

8、ew and approve protocol審核并批準驗證方案ü Assure all the equipment used for validation can be maintained timely保證驗證用所有設備均能按SOP進行及時的維護和保養ü Assure all the raw materials used for validation can be supplied timely and comply with domestic quality criteria of enterprise保證驗證用原輔包裝材料能按時到貨，并符合企業內控標準n Resea

9、rch and development department 研發部ü Provide product registration criteria and review whether recipe and process step described in the protocol is complying with registered criteria提供產品的注冊工藝，審核方案中生產處方、工藝與注冊資料的一致性ü Review and make assurance the compliance of raw material supplier with regist

10、ered supplier審核原輔料供應商和注冊資料的一致性ü Review relevant quality criteria including raw material, intermedia and finished product審核原輔料標準、中控質量標準和成品的質量標準n Vaccine one Plant 疫苗一室ü Coordination and implementation of this validation protocol協調、實施本驗證方案ü Training of personnel based on this validation

11、 protocol按驗證方案對有關人員進行培訓ü Collection, sorting and reviewing of validation data and drafting of validation reports (including the deviation report, if there are any deviations)收集、整理和審核驗證數據，起草驗證報告（如方案執行有偏差，要完成偏差報告）。n Utility support department 工程服務部ü Assistance in implementation of this valid

12、ation protocol協助本驗證方案的實施ü Calibration of instruments related to this validation protocol and providing relevant calibration certificates and quality certificates負責校驗和本驗證方案有關的儀器、儀表，出具校驗報告并貼合格證ü Assure normal operation of the utility system保證公用系統的正常運行ü Final report Review 最終報告的審核n Quali

13、ty control of quality department 質量部質量控制ü Determination of test methods used in this validation protocol負責確定本驗證方案的檢驗方法ü Validation of the necessary test methods for XXXXX involved in this validation protocol負責本驗證方案涉及的重組乙型肝炎疫苗（CHO細胞）原液生產的必要的檢驗方法的驗證ü The quality inspections on XXXXX Tab

14、let involved in this validation protocol, and presenting full item certificates of analysis and quality inspection reports and records負責本驗證方案涉及的重組乙型肝炎疫苗（CHO細胞）原液生產的質量檢驗，并出具全項檢驗報告，提供質量檢驗報告書及檢驗記錄ü Providing technical supports and guidance for tests at each of the process steps負責各工序檢驗的技術支持和指導ü

15、; Coordinating the plant in the implementation of this validation protocol協同車間實施本驗證方案ü Final report Review 最終報告的審核n Quality assurance of quality department 質量部質量保證ü Supervision of validation process, review and approval of this protocol, and ensuring the implementation following this proto

16、col負責驗證過程監督，方案的審核與批準，確保按批準的方案執行ü Sampling at each process steps during the process validation負責工藝驗證中各工序的取樣ü Coordination among different departments involved in the validation activities負責驗證涉及到的各部門之間的協調ü Providing equipment validation reports related to this validation protocol負責提供和本驗

17、證方案有關的設備驗證報告ü Identification, audit and assessment of suppliers供應商的確認、審計及評價ü Assessment and handling of changes and deviations occur during this process validation負責對工藝驗證實施過程中出現的變更和偏差的評價和處理ü Final report Review 最終報告的審核4. Regulation and Guidance 法規和指南ü EudraLex Volume 4 EU Guideli

18、nes to Good Manufacturing Practice Medicinal Products for Human and Veterinary Use, November, 2008EudraLex 第四卷 歐盟藥品生產質量管理規范指南 醫藥產品 人用及獸用，2008年11月ü Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice : Qualification and Validation , September 2001EudraLex 第四卷 藥品生產質量管理規范，附錄15

19、 驗證與確認，2001年9月ü EMEA - Note for guidance on process validation, September 2001 歐洲藥品管理局 工藝驗證指南，2001年9月ü EMEA - Annex II to note for guidance on process validation, January 2005歐洲藥品管理局 附錄II 工藝驗證指南ü FDA 21 Code of Federal Regulations, Part 211, current Good Manufacturing Practice for Fin

20、ished Pharmaceuticals, April 2011FDA聯邦法典第21篇第211部分，現行藥物制劑生產質量管理規范，2011年4月ü FDA Process Validation: General Principles and Practices , January 2011FDA工藝驗證：一般原則和實踐，2011年01月ü SFDA Good Manufacturing Practice (2010 Revision), March, 2011SFDA 藥品生產質量管理規范（2010年修訂），2011年03月ü PIC PIC/S Pharmac

21、eutical Inspection Convention, Pharmaceutical Inspection Co-operation Scheme, “Recommendations on Validation Master Plan, Installation and Operational Qualification, Non-sterile Process Validation, Cleaning Validation”, July 2004PIC/S藥品監管公約，藥品監管合作計劃，“有關驗證主計劃、安裝和運行確認、非無菌工藝驗證、清潔驗證的建議”，2004年7月ü A

22、WHO Guide to good manufacturing practice(GMP) requirements , Part 2 Validation , 1997 WHO 關于GMP要求的指南， 第二部分：驗證，1997年ü Chinese Pharmacopoeia (CP) 2010 Edition 中國藥典（CP）2010年版ü Process Validation of Protein Manufacturing (Technical Report No. 42 Supplement Vol. 59, No. S-4 September/October 20

23、05 © 2005 by PDA)蛋白生產的工藝驗證（技術報告42，PDA， 2005年版）5. Abbreviations縮略語The abbreviations which will be used in this protocol are listed in the following form.在下面的表格中規定了本方案中使用的縮略語。表格 1 縮略語Abbreviations縮略語Definition定義APIActive Pharmaceutical Ingredient (or “Drug Substance”)原料藥（或藥品物質）CPPCritical Process

24、 Parameter關鍵工藝參數CQACritical Quality attribute關鍵質量屬性CVCleaning Validation清洗驗證IQInstallation Qualification安裝確認MFVMedia Fill Validation培養基模擬灌裝試驗N/ANot applicable不適用OQOperation Qualification運行確認PLCProgrammable Logic Controller可編程邏輯控制器PQPerformance Qualification性能確認PVProcess Validation工藝驗證PWPurified Wate

25、r純化水QAQuality Assurance質量保證RARisk Assessment風險分析SOPStandard Operation Procedure標準操作程序URSUser Requirement Specification用戶使用要求VMPValidation Master Plan驗證主計劃WFIWater for Injection注射用水6. Protocol Description方案說明n Using this defined process validation shall be performed for the results of the Process Ris

26、k Assessment.本方案實施的工藝驗證，是基于工藝風險評估結果的。n Using this defined process validation shall be performed for three consecutive successful runs based on GMP requirements to demonstrate the process control reliability and the repeatability.鑒于XXX的生產已經有近20年歷史，本次驗證僅是對生產地址發生變更后且主要設備未發生變化的再驗證，考慮到本產品生產周期長（總生產周期為158天

27、）的原因，本方案實施的工藝驗證將生產1批成功的產品以證明其可靠性。n The determination of the intermediate and final product will implement enterprise internal standard but release finished product should be in accordance with criteria of the item “XXX” in Chinese Pharmacopoeia 2010 Edition.n 半成品、成品按企業內控標準檢驗，成品發放按中國藥典2010版三部“XXX ”標準

28、執行Raw material internal standard原輔料內控標準：表格 2原輔料質量標準表Quality Standard Doc.質量控制標準文件Doc. Number文件編號Sodium Chloride (BP) Quality Specification氯化鈉（藥用級）檢驗質量標準Sodium Dihydrogen Phosphate (BP) Quality Specification磷酸二氫鈉（藥用級）檢驗質量標準Crystal Aluminum Chloride AR Quality Specification 結晶氯化鋁AR檢驗質量標準Glucose (BP) Q

29、uality Specification 葡萄糖（藥用級）檢驗質量標準Glutamine Quality Specification 谷氨酰胺質量標準Potassium Bromide Quality Specification 溴化鉀質量標準Disodium Edetate Quality Specification 乙二胺四乙酸二鈉質量標準Sodium Hydroxide (BP) Quality Specification 氫氧化鈉（藥用級）質量標準 Disodium Hydrogen Phosphate (BP) Quality Specification 磷酸氫二鈉(藥用級)質量標準

30、Merthiolate (BP) Quality Specification 硫柳汞（藥用）質量標準Hydrochloric Acid (BP) Quality Specification 鹽酸（藥用級）檢驗質量標準Proline (BP) Quality Specification 脯氨酸（藥用）質量標準Sodium Bicarbonate (BP) Quality Specification 碳酸氫鈉（藥用級）檢驗質量標準DMEM Quality Specification DMEM質量標準Formaldehyde Solution (BP) Quality Specification 甲

31、醛溶液(藥用)質量標準Ammonium Sulfate AR Quality Specification 硫酸銨AR質量標準 Glycine (BP) Quality Specification 甘氨酸（藥用）質量標準Trypsin (BP) Quality Specification 胰蛋白酶(藥用)質量標準 New Born Bovine Serum Quality Specification 新生牛血清質量標準Methotrexate氨甲喋呤Dimethyl Sulphoxide二甲基亞砜Semi-product of XXX standard, include the Harvest

32、liquid, the Purification liquid and the Bulk.XXX中間品質量標準，包括細胞收獲液、純化產物、原液（文件號：XXX）Final product of XXX standard XXX成品質量標準：（文件號：XXX）n The validation procedures, items and contents are divided based on the system process steps. The process steps, validation items, sampling methods, testing methods and a

33、cceptance criteria are described for each of the process steps.驗證的程序、項目和內容中以系統工序為單位，對各工序中的所涉及的工藝步驟、驗證項目、取樣方法、檢驗方法和接受標準進行了規定和描述。n The process recipe of Bulk of XXX (Registration batch No.: for 20g: GUOYAOZHUNZI XXX, and for 10g GUOYAOZHUNZI XXX) is briefly described in this protocol. The contents of

34、the recipe are described and the process flow diagram is listed. The process procedure is conformed to the registration standard.本方案對XXX原液的工藝處方（注冊批號：20g：國藥準字 XXX、10g：國藥準字 XXX）進行了簡介，對處方內容進行描述，并且列出直觀化的工藝流程圖。工藝規程與注冊標準一致。n This protocol is used to specify the purpose and the scope of this validation and

35、 the responsibilities of each of the departments during the implementation of this validation protocol.本方案旨在指明驗證的目的、范圍，并規定了該驗證方案在實施過程中的各部門的職責。n Summarize and evaluate the entire process validation based on the validation of each of the process steps and record the evaluation results to the report.在各

36、工序驗證的基礎上，對整個工藝驗證進行了總結評價，評價結果記錄到驗證報告中。7. Product And Process Description產品和工藝描述7.1 Product specification(1) Ampoule, one human dose of 1.0ml, containing 10g of HBsAg (1) 安瓿瓶，每1次人用劑量為1.0ml，含HBsAg 10g；(2) Ampoule, one human dose of 1.0ml, containing 20g of HBsAg (2) 安瓿瓶，每1次人用劑量為1.0ml，含HBsAg 20g；(3) Via

37、l, one human dose of 1.0ml, containing 20g of HBsAg (3) 西林瓶，每1次人用劑量為1.0ml，含HBsAg 20g；(4) Vial, one human dose of 0.5ml, containing 10g of HBsAg (4) 西林瓶，每1次人用劑量為0.5ml，含HBsAg 10g。7.2 Drug approval No. 20g：GUOYAOZHUNZI S20g：國藥準字 S10g：GUOYAOZHUNZI S10g：國藥準字 S7.3 Standard Followed: Chinese Pharmacopoeia

38、2010 Edition執行標準：中國藥典2010版7.4 Raw Materials and Adjuvants原輔料情況Table 3 Raw Materials and Adjuvants表格 4原輔料信息表Name SpecificationsManufacturerPackaging SpecificationsSodium Chloride BP Tianjin Haiguang Pharmaceutical Industrial Co., Ltd. 1000g/bagDisodium hydrogen phosphate BP Hunan Jiudian Pharmaceutic

39、al Co., Ltd. 0.5kg/bottleCrystal Aluminum Chloride AP Beijing Yili Fine Chemical Co., Ltd. 500g/bottleGlucose AP Baitaike Bioltechnology Co., Ltd. (filling)500g/bottleGlutamineusp Gradeinvitrogen corporation (Gibco)1kg /drum Potassium Bromide AP Beijing Yili Fine Chemical Co., Ltd. 500g/bottleDisodi

40、um Edetate AP Sinopharm Chemical Reagent Co., Ltd 250g/bottleSodium Hydroxide AP Beijing Chemical Factory 500g/bottleMono-water disodium hydrogen phosphate BP Hunan Jiudian Pharmaceutical Co., Ltd. 0.5kg/bottleMerthiolate BP (USP/BP/EP)GIHON1kg /drum Hydrochloric acid AP Beijing Chemical Factory 250

41、0ml/ bottleProline -Shanghai Xiehe Amino Acid Co., Ltd. 500g/bagSodium bicarbonate AP Beijing Chemical Factory 500g/bottleDMEM-Gibco6687g/drum Formaldehyde AP Sinopharm Chemical Reagent Co., Ltd 500ml/ bottleAmmonium sulfate AP Beijing Yili Fine Chemical Co., Ltd. 25kg /drum GlycineAP Sinopharm Chem

42、ical Reagent Co., Ltd 100g/ bottle Pancreatin -BD 500g/bottleGibco100g/ bottle Methotrexate99.0%sigma-aldrich（fluka-analytical）100mg/ tube Dimethyl Sulphoxide-SERVA Electrophoresis GmbH1L/ bottle New born bovine serum Top grade Wuhan Sanli Biotechnology Co., Ltd. 1000ml/ bottle-Inner Mongolia Jinyua

43、nkang Biotechnology Co., Ltd. 500ml/ bottle原輔料名稱規格生產廠家包裝規格氯化鈉藥用級天津海光藥業有限公司1000g/袋磷酸氫二鈉藥用級湖南九典制藥有限公司0.5kg/瓶結晶氯化鋁分析純北京益利精細化學品有限公司500g/瓶葡萄糖分析純寶泰克生物科技公司（分裝）500g/瓶谷氨酰胺usp Gradeinvitrogen corporation（Gibco）1kg/桶溴化鉀分析純北京益利精細化學品有限公司500g/瓶乙二胺四乙酸二鈉分析純國藥集團化學試劑有限公司250g/瓶氫氧化鈉分析純北京化工廠500g/瓶一水磷酸二氫鈉藥用級湖南九典制藥有限公司0.5

44、kg/瓶硫柳汞藥用級（USP/BP/EP）GIHON1Kg/桶鹽酸分析純北京化工廠2500ml/瓶脯氨酸-上海協和氨基酸有限公司500g/袋碳酸氫鈉分析純北京化工廠500g/瓶DMEM-Gibco6687g/桶甲醛分析純國藥集團化學試劑有限公司500ml/瓶硫酸銨分析純北京益利精細化學品有限公司25kg/桶甘氨酸分析純國藥集團化學試劑有限公司100g/瓶胰酶-BD公司500g/瓶Gibco100g/瓶氨甲喋呤99.0%sigma-aldrich（fluka-analytical）100mg/管二甲基亞砜-SERVA Electrophoresis GmbH1L/瓶新生牛血清優級武漢三利生物技術

45、有限公司1000ml/瓶-內蒙古金源康生物工程有限公司500ml/瓶7.5 Process description:工藝簡述：Ø Process Cycle 工藝周期Table 5 Process Schedule表格 6工藝時間表No.序號Process Steps 工藝步驟Time (Day)時間（天）1Resurrection 復蘇12C25 - First square bottle incubation C25-方瓶第一次培養573C26 - second square bottle incubation C26-方瓶第二次培養574C27 - Kolle flask in

46、cubation C27-克氏瓶培養575C28 - First 3L rotary bottle incubation C28-3L轉瓶第一次培養576C29 - second 3L rotary bottle incubation C29-3L轉瓶第二次培養577C30 - third 3L rotary bottle incubation C30-3L轉瓶第三次培養578C31 - first 15L bottle incubation C31-15L瓶第一次培養579C32 - second 15L bottle incubation C32-15L瓶第二次培養571020 conse

47、cutive harvest 連續20次收獲液4011Filtration for cell residue removal 去細胞殘渣過濾0.512UF concentration 超濾濃縮0.513Ammonium sulfate precipitation 硫酸銨沉淀4514Potassium bromide extraction 溴化鉀提取115First density gradient centrifugation 一次密度梯度離心1.516Second density gradient centrifugation 二次密度梯度離心1.517UF concentration 超濾

48、濃縮118Chromatography purification 層析純化119Bulk solution combination 原液合并0.5Ø Preparation of production cells生產用細胞制備Take 1 to 2 tubes of cells from the working cell bank (of a same batch No.), resurrect, mix and inoculate them into a 100ml cell cultivation bottle in a ratio of 1:1. After the cells

49、 have grown full and adhering to the wall to a single layer, perform the 0.25% trypsin digestion and passage. And then inoculate them into a 100ml cell cultivation bottle in a ratio of 1:4. Passage and proliferate following the sequence of 100ml cell cultivation bottle, Kolle flask and the rotary bo

50、ttle. The ratio is determined according to different cell cultivation bottle specifications. Two 100ml cell cultivation bottles are used for the passage to 1 Kolle flask. One Kolle flask is used for the passage to 1 3L rotary bottle. One 3L rotary bottle is used for the passage to 7 to 8 3L rotary b

51、ottles. One 3L rotary bottle is used for the passage to 1 15L rotary bottle. One 15L rotary bottle is used for the passage to 7 to 8 15L rotary bottles. Incubate them while the bottles are kept still or rotated at 36±1 (the 100ml cell cultivation bottles are kept still during incubation and rot

52、ary bottles are rotated). The duration of each expansion incubation step is 5 to 7 days. After proliferated to generation No. 32, the change solution is maintained. 取工作細胞庫細胞12支（同一批號），復蘇混合后按1：1比例接種100ml細胞培養瓶，待細胞貼壁長滿至單層后，0.25%胰蛋白酶消化傳代，按1：4比例接種100ml細胞培養瓶。按100ml細胞培養瓶克氏瓶轉瓶傳代擴增，比例根據不同規格細胞培養瓶而定，2個100ml細胞培養

53、瓶傳1個克氏瓶；1個克氏瓶傳1個3L轉瓶；一個3L轉瓶傳78個3L轉瓶；一個3L轉瓶傳1個15L轉瓶；1個15L轉瓶傳78個15L轉瓶。置36±1條件下，采用靜止或旋轉培養（100ml細胞培養瓶和克氏瓶采用靜止培養，轉瓶采用旋轉培養法）。每次擴增培養時間為57天，傳代擴增至32代，開始維持換液。 Ø Incubation media培養液The growth media used for cell passage and proliferation is DMEM solution containing 9% to 10% inactivated new born bovi

54、ne serum with a final concentration of methotrexate of 10-6mol/L and L-glutamine of 2mmol/L. The maintenance solution is DMEM solution containing 3% to 5% inactivated new born bovine serum. 細胞傳代擴增用生長液為含有9%10%滅能新生牛血清的DMEM液，其中含終濃度為10-6mol/L氨甲喋呤、2mmol/L L-谷氨酰胺。維持液為含有3%5%滅能新生牛血清的DMEM液。Ø Harvesting收

55、獲After passage to the No. 32 generation and the cells have grown into a compact single layer adhering to the wall (Day 5 to 7), discard the growth liquor, add the maintenance solution, discard the incubation liquor 48 hours later and add the maintenance solution. Replace the maintenance solution every 48 hours thereafter and harvest the incubation liquor. Measure the HBsAg content in the harvested supernatant with ELISA (or reverse hemagglutination method), the result shall be above 1.0mg/L