【正文】 Personal Identification人員確認(rèn) 51 Process Documentation Verification 生產(chǎn)文件確認(rèn) 51 Training Verification 培訓(xùn)確認(rèn) 52 System and Equipment Instruments Calibration Verification 系統(tǒng)和設(shè)備儀表校驗(yàn)確認(rèn) 52 System and Equipment Instruments Calibration Verification 生產(chǎn)相關(guān)驗(yàn)證狀態(tài)確認(rèn) 53 Test Instruments Calibration Verification 測(cè)試儀器和儀表校驗(yàn)確認(rèn) 53 Testing Method Validation Verification檢驗(yàn)方法驗(yàn)證的確認(rèn) 54 Raw Material and Excipient Verification原輔料確認(rèn) 549. Sampling Plan and Evaluation Criteria取樣計(jì)劃及評(píng)估標(biāo)準(zhǔn) 5610. Sample Coding Method樣品編碼原則 7011. Process Validation Test Execution 工藝驗(yàn)證執(zhí)行 72 Liquid Preparation 液體配制 72 Cell Resurrection and Passage細(xì)胞復(fù)蘇及傳代 74 Harvest liquid test 收獲液檢測(cè) 81 Clarification Filtration Efficacy Test 澄清過(guò)濾效果檢測(cè) 83 UF Concentration 1 Efficacy Test超濾濃縮1效果檢測(cè) 83 Ammonium sulfate precipitation efficacy test 硫酸銨沉淀效果檢測(cè) 85 Potassium bromide extraction efficacy test 溴化鉀提取效果檢測(cè) 86 First density gradient centrifugation efficacy test 一次密度梯度離心效果檢測(cè) 87 Second density gradient centrifugation efficacy test 二次密度梯度離心效果檢測(cè) 88 UF Concentration 2 Efficacy Test 超濾濃縮2效果檢測(cè) 89 Chromatography purification product test 層析純化產(chǎn)物檢測(cè) 9012. Bulk Determination原液檢驗(yàn) 9213. Process Validation Summarization工藝驗(yàn)證總結(jié) 9314. PV Report工藝驗(yàn)證報(bào)告 94 Process Validation Report 工藝驗(yàn)證報(bào)告 94 Process Validation Analysis and Evaluation 工藝驗(yàn)證分析與評(píng)價(jià) 9415. Requirements on Execution Records 執(zhí)行記錄要求 9516. Deviation Handling 偏差處理 9517. Change Handling 變更處理 9518. Test Form測(cè)試表 96表格 1 縮略語(yǔ) 9表格 2原輔料質(zhì)量標(biāo)準(zhǔn)表 11表格 3物料消耗表1 21表格 4物料消耗表2 21表格 5所用液體配料表 22表格 6生產(chǎn)車間一覽表 26表格 7主要生產(chǎn)設(shè)備一覽表 29表格 8工藝風(fēng)險(xiǎn)分析表 33表格 9工藝時(shí)間表 49表格 10取樣計(jì)劃表 56表格 11樣品編碼原則 70表格 12配液取樣檢測(cè)信息表 72表格 13復(fù)蘇和傳代取樣表 74表格 14復(fù)蘇和傳代樣品編碼 80表格 15收獲液標(biāo)準(zhǔn) 82表格 16澄清過(guò)濾標(biāo)準(zhǔn) 83表格 17超濾濃縮1標(biāo)準(zhǔn) 84表格 18硫酸銨沉淀標(biāo)準(zhǔn) 85表格 19溴化鉀提取標(biāo)準(zhǔn) 86表格 20一次密度梯度離心標(biāo)準(zhǔn) 87表格 21二次密度梯度離心標(biāo)準(zhǔn) 88表格 22超濾濃縮2標(biāo)準(zhǔn) 89表格 23純化產(chǎn)物標(biāo)準(zhǔn) 90表格 24原液標(biāo)準(zhǔn) 92表格 25測(cè)試表列表 961. Purpose目的This purpose of this process validation is to provide documented evidence to show that the bination 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 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(yàn)證用于提供文件化的證據(jù)，證明XXX疫苗一室生產(chǎn)XXX原液的人員、材料、設(shè)備、方法、環(huán)境條件以及其它有關(guān)公用設(shè)施的組合可以始終如一的生產(chǎn)出符合企業(yè)內(nèi)控標(biāo)準(zhǔn)及國(guó)家法定標(biāo)準(zhǔn)的產(chǎn)品，工藝穩(wěn)定可靠，符合GMP要求，工藝驗(yàn)證的過(guò)程和檢查的結(jié)果將按照該驗(yàn)證方案進(jìn)行記錄。 Assure all the equipment used for validation can be maintained timely保證驗(yàn)證用所有設(shè)備均能按SOP進(jìn)行及時(shí)的維護(hù)和保養(yǎng)252。 Providing technical supports and guidance for tests at each of the process steps負(fù)責(zé)各工序檢驗(yàn)的技術(shù)支持和指導(dǎo)252。 SFDA Good Manufacturing Practice (2010 Revision), March, 2011SFDA 藥品生產(chǎn)質(zhì)量管理規(guī)范（2010年修訂），2011年03月252。 Preparation of production cells生產(chǎn)用細(xì)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 have grown full and adhering to the wall to a single layer, perform the % 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 bottle. 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 bottles. 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 36177。 Purification純化u Filtration過(guò)濾The harvested culture media go first through a filter and then a filter to remove any cell residues. ，過(guò)濾去除細(xì)胞碎片。1℃保溫72小時(shí)。 Sterilization of Purified Product 純化產(chǎn)物除菌 After acceptable results are obtained, HBsAg purified products from a same cell digestion batch are bined. And the bulk is obtained after UF, concentration and sterile filtration. 同一細(xì)胞消化批來(lái)源的HBsAg純化產(chǎn)物檢定合格后合并，經(jīng)超濾、濃縮、。u Ammonium sulfate precipitation 硫酸銨沉淀After UF concentration, 312g of ammonium sulfate is added to each liter of the obtained culture media. It is then allowed to stand at room temperature for 4 to 5 days after stirring to allow HBsAg to precipitate. 超濾濃縮后的培養(yǎng)液每升加入312g硫酸銨，攪拌均勻后，室溫放置4～5天，使HBsAg沉淀。按100ml細(xì)胞培養(yǎng)瓶→克氏瓶→轉(zhuǎn)瓶傳代擴(kuò)增，比例根據(jù)不同規(guī)格細(xì)胞培養(yǎng)瓶而定，2個(gè)100ml細(xì)胞培養(yǎng)瓶傳1個(gè)克氏瓶；1個(gè)克氏瓶傳1個(gè)3L轉(zhuǎn)瓶；一個(gè)3L轉(zhuǎn)瓶傳7～8個(gè)3L轉(zhuǎn)瓶；一個(gè)3L轉(zhuǎn)瓶傳1個(gè)15L轉(zhuǎn)瓶；1個(gè)15L轉(zhuǎn)瓶傳7～8個(gè)15L轉(zhuǎn)瓶。 A WHO Guide to good manufacturing practice(GMP) requirements , Part 2 Validation , 1997 WHO 關(guān)于GMP要求的指南， 第二部分：驗(yàn)證，1997年252。 Final report Review 最終報(bào)告的審核n Quality assurance of quality department 質(zhì)量部質(zhì)量保證252。 Provide product registration criteria and review whether recipe and process step described in the protocol is plying with registered criteria提供產(chǎn)品的注冊(cè)工藝，審核方案中生產(chǎn)處方、工藝與注冊(cè)資料的一致性252。Protocol for XXXXXX工藝驗(yàn)證方案Doc. No.文件編號(hào)Page頁(yè)碼Process Validation (PV) Coordination and implementation of this validation protocol協(xié)調(diào)、實(shí)施本驗(yàn)證方案252。 Coordination among different departments involved in the validation activities負(fù)責(zé)驗(yàn)證涉及到的各部門(mén)之間的協(xié)調(diào)252。 2005 by PDA)蛋白生產(chǎn)的工藝驗(yàn)證（技術(shù)報(bào)告42，PDA， 2005年版）5. Abbreviations縮略語(yǔ)The abbreviations which will be used in this protocol are listed in the following form.在下面的表格中規(guī)定了本方案中使用的縮略語(yǔ)。每次擴(kuò)增培養(yǎng)時(shí)間為5～7天，傳代擴(kuò)增至32代，開(kāi)始維持換液。Inject the samples following the sequence of solution A, solution B, solution C and solution D. The speed of the centrifuge is 2000rpm both at the time of sample injection and at the time of sample collection. The centrifugation is performed at a speed o 25000rpm for 22 hours. HBsAg peaks are collected according to the UV absorbance value at 280nm. The HBsAg peak is peak B. The same method is used for the collection