WHO对HPV疫苗质量安全性及有效性指导原则
疫苗临床试验技术指导建议建议原则
研究参与者与合作方管理
研究者培训
对研究者进行培训，确保其了解试验方案和操作 规程。
合作方选择
选择具备资质和经验的合作方参与试验，确保试 验质量和合规性。
合同协议
与合作方签订明确的合同协议，明确双方的权利 和义务。
THANKS
感谢观看
盲法原则
为避免主观偏见对结果的影响，应采用盲法原则，如单盲、双盲等，确保试 验结果的客观性和准确性。
适应性临床试验
适应性临床试验
在临床试验过程中，根据初步数据分析结果，对试验方案进行适 时调整，以优化试验效率和质量。
适应性临床试验的优势
能够及时调整试验方案，提高试验效率和质量，减少不必要的浪 费和风险。
04
疫苗临床试验监控
数据监控和安全性评估
总结词：严格把控
详细描述：在疫苗临床试验过程中，需要对试验数据进行实时监控，确保数据的准确性和完整性。同 时，对疫苗的安全性进行评估，包括观察受试者是否有过敏反应或其他不良反应。
不良事件和严重不良事件报告
总结词
及时准确报告
VS
详细描述
在疫苗临床试验过程中，一旦发现受试者 出现不良事件或严重不良事件，应立即报 告给相关机构和伦理委员会，以便及时采 取相应措施。
适应性临床试验的挑战
需要严格控制试验过程和数据分析，确保试验的可靠性和稳定性 。
03
疫苗临床试验实施
试验机构和人员要求
试验机构
应具备完善的组织架构和健全的实验室条件，能够承担疫苗临床试验的所有环节 。
人员要求
试验人员应具备相关的教育背景和丰富的实践经验，并接受过系统的培训和考核 ，具备疫苗临床试验的专业知识和技能。
分析结果等。
HPV疫苗效果及安全性评估
HPV疫苗效果及安全性评估《HPV 疫苗效果及安全性评估》在当今的医疗领域，HPV 疫苗成为了备受关注的焦点。
HPV 即人乳头瘤病毒，是一种常见的性传播病毒，与多种癌症的发生密切相关，尤其是宫颈癌。
HPV 疫苗的出现为预防 HPV 感染和相关疾病带来了新的希望。
但对于大众来说，了解HPV 疫苗的效果以及安全性至关重要。
首先，让我们来看看 HPV 疫苗的效果。
大量的临床研究和实际应用数据表明，HPV 疫苗在预防 HPV 感染方面具有显著的效果。
HPV 疫苗可以预防多种 HPV 型别，包括一些高危型别，如 HPV16 和 HPV18，这两种型别是导致宫颈癌的主要“元凶”。
通过接种疫苗，人体可以产生特异性的抗体，这些抗体能够在 HPV 病毒入侵时迅速做出反应，将其清除，从而降低感染的风险。
对于未感染过 HPV 的人群，尤其是青少年和年轻女性，接种 HPV疫苗能提供近乎 100%的预防特定型别 HPV 感染的保护作用。
这意味着，如果在首次性生活之前接种疫苗，能够最大程度地发挥疫苗的预防效果，显著降低未来患宫颈癌等疾病的风险。
除了对个体的保护作用，HPV 疫苗还具有群体免疫的效果。
当足够多的人群接种了疫苗，病毒在人群中的传播就会受到抑制，从而间接保护了那些因各种原因未能接种疫苗的人群。
然而，HPV 疫苗的效果并非是一劳永逸的。
其保护作用可能会随着时间的推移而有所减弱，但现有研究表明，接种后的长期保护效果依然可观。
而且，定期的宫颈癌筛查仍然是必不可少的，即使接种了疫苗，也不能完全替代筛查的重要性。
接下来，我们探讨一下 HPV 疫苗的安全性。
HPV 疫苗在上市前经过了严格的临床试验和审批程序，其安全性得到了广泛的评估。
常见的不良反应通常是轻微和短暂的，比如注射部位的疼痛、红肿、发热、头痛等，这些症状一般在接种后的几天内自行消失，不会对身体造成长期的不良影响。
严重的不良反应极为罕见。
从全球范围内的大规模接种情况来看，HPV 疫苗的安全性是有保障的。
WHO非传染性疾病预防控制监测框架和目标
世界卫生组织讨论文件（修订版）(2012年7月25日版)全球非传染性疾病预防控制综合监测框架（含指标）和自愿性目标前言非传染性疾病（NCDs）位列全球死因之首。
2008年，全球有5700万人死亡，其中3600万人死于非传染性疾病（主要包括心血管疾病、癌症、糖尿病和慢性肺部疾病），几乎占全部死因的2/3(1)。
由上述疾病所致负担在低收入国家中正快速增长。
全球大约1/4的非传染性疾病死亡发生在60岁以前(2)。
大部分非传染性疾病是可以预防的。
它们有共同的、可改变的行为危险因素，如烟草使用、不健康膳食、身体活动不足和有害饮酒。
这些危险因素导致超重和肥胖、血压升高以及胆固醇升高。
如果不采取行动，在未来的30年，非传染性疾病负担所造成的花费将使数万亿美元的资源付之东流(3)。
目前已有了一些切实可行且经济有效的干预措施，以降低非传染性疾病的负担和影响，而持续不懈地预防危险因素和改善卫生保健将可避免数百万人过早死亡(4)。
《全球预防和控制非传染性疾病战略行动计划》(5)提出以下关键要素：监测、预防和卫生保健。
监测的目的是监视非传染性疾病并分析其在社会、经济、行为及政治方面的决定因素，以便为政策、法律和财政措施的制定提供指导。
2011年10月19日至20日，联合国在美国纽约举行了“预防和控制非传染性疾病高级别会议”，会议强调了监测和监督非传染性疾病预防和控制进展的重要性。
2011年10月19日联大会议通过了“关于预防和控制非传染性疾病问题高级别会议的政治宣言”的第66/2号决议(6)。
决议要求世卫组织（WHO）在2012年底之前完成以下工作：i.制定全球综合监测框架，包括一套可适应于不同区域和国家的指标，还包括通过多部门合作监测非传染性疾病趋势，并评估在实施国家非传染性疾病预防控制策略和计划方面所取得的进展。
ii.就全球非传染性疾病预防控制自愿性目标提出建议。
联大会议政治宣言也敦促其成员国根据国情，并以世卫组织的指导为基础，考虑和制定本国的目标和指标。
生物制药在疫苗生产中的质量控制标准与监管考核试卷
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三、填空题（本题共10小题，每小题2分，共20分，请将正确答案填到题目空白处）
6.疫苗的免疫接种通常分为______剂次。
7.疫苗生产过程中，用于去除病毒和支原体的常用方法是______。
8.下列哪种技术常用于疫苗的冷冻干燥处理？______
9.疫苗生产中，用于检测疫苗效力的常用方法是______。
10.在我国，负责疫苗上市审批的机构是______。
四、判断题（本题共10小题，每题1分，共10分，正确的请在答题括号中画√，错误的画×）
D.疫苗的不良反应监测
4.在疫苗生产过程中，以下哪种技术常用于分离纯化疫苗成分？（）
A.电泳技术
B.色谱技术
C.显微镜技术
D. X射线技术
5.以下哪种疫苗属于重组蛋白疫苗？（）
A.百白破疫苗
B.肺炎疫苗
C. HPV疫苗
D.麻疹疫苗
6.在疫苗生产过程中，以下哪个环节最容易产生批次间的差异？（）
A.原材料质量控制
1. C
2. A
3. C
4. B
5. C
6. B
7. A
8. D
9. B
10. A
11. B
12. C
13. C
14. C
15. D
16. A
HPV疫苗的申报现状及药学研发思考
HPV疫苗的申报现状及药学研发思考摘要：自2006年世界上首支人乳头瘤病毒(HPV) 疫苗注册上市以来，多家国内企业开始了 HPV 疫苗的研发及申报工作。
本文在简要介绍HPV疫苗申报现状的基础上，重点讨论了HPV申报资料中的药学问题，并提出了相应的建议，以期起到抛砖引玉之效。
关键词：HPV疫苗；申报现状；药学研发HPV疫苗的研发与上市让人们能够有效预防人乳头瘤病毒感染，因而引起了各个国家与社会各界的广泛关注。
目前，我国HPV疫苗的研发虽取得了一定的成果，但仍需要进一步提高研发效率与申报质量，一、HPV疫苗申报现状（一）已上市的HPV疫苗就现阶段而言，国际上已有3种 HPV 疫苗获准上市使用，分别为2价疫苗(HPV16/18)、4价疫苗(HPV6/11/16/18)及9价疫苗(HPV6/11/16/18/31/33/45/52/58) ，以上产品均于近年在中国获准上市[1]。
2019年12月，我国企业所研发的2价HPV疫苗( HPV16 /18) 取得了上市资格。
（二）已获批及处于申请阶段的HPV疫苗随着对人乳头瘤病毒的研究愈加深入，近年来HPV疫苗的研发也得到了较好的发展，目前已有15个HPV疫苗的临床试验申请获得批准。
HPV疫苗申请数量与获批数量的增加，也使得HPV疫苗在表达体系、纯化工艺、佐剂类型等方面产生了一定的差异，更具针对性。
另外，相关研发企业也日益成熟，能够研发不同价数的HPV疫苗。
二、HPV申报资料中的药学问题及建议（一）工程菌（1）从已申报的HPV疫苗来看，不同研发企业在氨基酸序列的选择上存在差异，包括其来源、序列等多个方面，申报企业需要对此进行规范的说明，并与已上市的HPV疫苗作对比，从而体现出该HPV疫苗可能对免疫原性的影响。
需要注意的是，如果存在目的基因改造的情况，那么还要说明该项改造对病毒样颗粒结构的影响。
在此前提下，当目的基因又存在 N 端/C 端截短情况，那么研发企业需要通过提供相应的依据与数据对全长序列产品、截短序列的质量特性比较等方面进行说明。
WHO对HPV疫苗质量、安全性及有效性指导原则
GUIDELINES TO ASSURE THE QUALITY, SAFETY AND EFFICACY OF RECOMBINANT HUMAN PAPILLOMA VIRUS VIRUS?LIKE PARTICLE V ACCINESHPV二、Special consideration section:在生产、非临床及临床中过程中的考虑因素:1、生产方面:VLP是复杂的生物产物,必须在不同水平下对其进行检测分析。
因而在其生产过程及质量控制上必须考虑以下几个因素:1)新的表达体系如杆状病毒(GSK),新的特殊要求。
但我们用的毕赤酵母,相对比较常见。
2)新佐剂(略)3)天然的L1蛋白是没有被糖基化修饰,目前的两种表达体系,在糖基化修饰上不存大问题,但要对糖基化及其位点进行分析。
4)L1衣壳蛋白亚单位的解聚与再聚,可能有利于纯化,并得到更稳定的VLP。
目前,我们的路线可能是不经解聚,直接纯化获得VLP。
个人感觉,到后期可以兵分两路,一路直接获得VLP,而另一路则将VLP解聚后,再进行纯化与重组。
5)纯化后的L1 VLP要进行生化及免疫上的鉴定,并测定L1的浓度、纯度及组聚情况。
6)如加入了防腐剂,应对其免疫性进行验证,并确认不会有负作用2、非临床方面:关键就是要证明其免疫原性,并能否产生免疫中和抗体。
3、临床方面:(略)三、生产指导(Part A. Guidelines on manufacturing)3.1定义definitions3.1.1国际名称和专有名称国际名称:重组人乳头瘤类病毒颗粒疫苗(基因型16 L1蛋白)3.1.2定义描述重组HPV VLP疫苗为无菌的液态疫苗,里面含纯化后由一种或多种HPV基因型重组的主要的衣壳蛋白,并与相应佐剂混合。
3.1.3国际标准品在此指导原则编写时,市场上暂无国际标准品提供。
但有相应试剂在实验室水平上,在进行注射后进行生物效价方面的评价如抗体滴度和病毒DNA检测。
WHO对HPV疫苗的指导原则
WHO/BS/06.2050 - FinalENGLISH ONLYEXPERT COMMITTEE ON BIOLOGICAL STANDARDIZATIONGeneva, 23 to 27 October 2006GUIDELINES TO ASSURE THE QUALITY, SAFETY ANDEFFICACY OF RECOMBINANT HUMAN PAPILLOMAVIRUSVIRUS-LIKE PARTICLE VACCINES© World Health Organization 2006All rights reserved. Publications of the World Health Organization can be obtained from WHO Press, World Health Organization, 20 Avenue Appia, 1211 Geneva 27, Switzerland (tel.: +41 22 791 3264; fax: +41 22 791 4857; e-mail: bookorders@who.int). Requests for permission to reproduce or translate WHO publications – whether for sale or for noncommercial distribution – should be addressed to WHO Press, at the above address (fax: +41 22 791 4806; e-mail: permissions@who.int).The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement.The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsed or recommended by the World Health Organization in preference to others of a similar nature that are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguished by initial capital letters.All reasonable precautions have been taken by the World Health Organization to verify the information contained in this publication. However, the published material is being distributed without warranty of any kind, either expressed or implied. The responsibility for the interpretation and use of the material lies with the reader. In no event shall the World Health Organization be liable for damages arising from its use. The named authors [editors] alone are responsible for the views expressed in this publication.Adopted by the 57th meeting of the WHO Expert Committee on Biological Standardization, 23-27 October2006. A definitive version of this document, which will differ from this version in editorial but not scientificdetails, will be published in the WHO Technical Report Series.WHO/BS/06.2050 - FinPage 2IntroductionGeneral considerationsSpecial considerationsPart A. Guidelines on manufacturingA.1DefinitionsA.2 General manufacturing recommendationsA.3Control of source materialsA.4Control of HPV antigen vaccine productionA.5Control of purified monovalent antigen bulkA.6Adsorbed monovalent antigen bulkA.7Final vaccine bulkA.8 Filling and containersA.9 Control tests on final vaccine lotA.10 RecordsA.11 Retained samplesA.12 LabellingA.13 Distribution and transportA.14 Stability testing, storage and expiry datePart B. Nonclinical evaluation of recombinant HPV VLP vaccinesB.1Pharmacological studiesB.2Safety Pharmacology studiesB.3Toxicology studiesPart C. Clinical evaluation of recombinant HPV VLP vaccinesC.1 Immune responses to the vaccineC.2 Studies of protective efficacyC.3 Bridging efficacy by means of immunogenicity dataC.4 Vaccine safetyC.5 Post-marketing studies and surveillancePart D. Guidelines for national regulatory authoritiesD.1GeneralD.2 Release and certificationAuthorsAcknowledgementsReferencesAppendix 1Model summary protocol for manufacturing and control of recombinant human papillomavirus virus-like particle vaccineAppendix 2Model certificate for the release of recombinant human papillomavirus virus-like particle vaccineWHO/BS/06.2050 - FinPage 3 IntroductionWHO convened two meetings in Geneva, 23-24 March and 28 - 30 August, 2006, where scientific experts, regulatory professionals and other stakeholders met todevelop guidelines for prophylactic human papillomavirus (HPV) vaccines. Thisdocument is intended to provide background and guidance to national regulatoryauthorities (NRAs) and vaccine manufacturers on the production, quality controland evaluation of the safety and efficacy of recombinant HPV virus-like particle(VLP) vaccines.This document sets out the guidance on product manufacture and qualityassessment in part A. In addition, guidance specific to the nonclinical and clinical evaluation of recombinant HPV vaccines is provided in Part B and Part C,respectively. This document should be read in conjunction with all relevant WHO guidelines including those on nonclinical (1) and clinical evaluation (2) ofvaccines. The following text is written in the form of guidelines instead ofrecommendations. Guidelines allow greater flexibility than recommendations with respect to expected future developments in the field. This guidance is based onthe experience of the products developed so far, as described below, and mayneed to be updated in view of future developments.General considerationsHPV is a small, non-enveloped deoxyribonucleic acid (DNA) virus. The circular, double-stranded viral genome is approximately 8-kb in length. The genomeencodes for 6 early proteins responsible for virus replication and 2 late proteins,L1 and L2, which are the viral structural proteins. L1 is the major structuralprotein. L1 proteins associate to form pentameric structures called capsomers.Mature virus particles are comprised of 72 capsomers arranged in icosahedralsymmetry. The minor capsid protein, L2, is present in as many as 72 moleculesper mature virus particle. L2 is not required for particle formation. HPVinfection, replication and particle maturation occurs in the stratified squamousepithelia of skin and mucous membranes, with virus spread occurring by skin-to-skin contact.Over 100 different types of HPV have been identified and molecularlycharacterized. These HPVs cause a variety of diseases in humans ranging frombenign warts to cancer of the epithelia (including the cervix, vagina, vulva, anusand oropharynx). Those HPV types associated with the development of cancerare called high risk for oncogenicity. Other HPV types, such as HPV types 6 and11 associated with genital warts, are considered low risk for oncogenicity.The majority of HPV infections by both high and low risk types are oftenasymptomatic, self-limiting and resolve spontaneously, presumably due to thehost immune response. In some instances, persistent infection by the high risktypes may ultimately progress to invasive carcinoma at the site of infection,WHO/BS/06.2050 - FinPage 4mainly of the genital tract, if not detected and treated appropriately. The interval between the acquisition of HPV infection and malignant progression usually takes about 10 years or longer. High risk HPV types can be detected in virtually allcases of cervical cancer, and it is generally accepted that the persistent viralinfection is necessary for the development of cancer (3). The basis forprogression to invasive carcinoma is not well defined. However, environmentaland physiological co-factors may increase the risk for cancer development inpersistently infected persons.The International Agency for Research on Cancer (IARC) has currently definedthirteen high risk HPV types that are associated with cancers in humans (4).Distribution and prevalence of these HPV types in cancer cases is generallyconsistent around the world. Two of the high risk HPV types, 16 and 18, account for approximately 70% of all cervical cancers globally (4). Most other genitalcancers, such as cancers of the vagina and anus are also associated with persistent HPV infection. In addition, these HPVs are associated with a fraction of cancers of the vulva, penis, and oropharynx. The incidence of cervical cancer issignificantly higher than all other HPV related cancers, and is the second mostcommon cancer among women worldwide.Low risk HPV types cause genital warts, recurrent respiratory papillomatosis(RRP), and low grade cervical dysplasia. The lifetime risk of genital wartsexceeds 10%. While not malignant, these lesions are associated with physical and psychological morbidity. They are also difficult to treat. RRP is a devastating,albeit rare, disease that manifests as recurrent, rapidly growing benign laryngealtumors that require frequent excision to prevent airway obstruction. HPV 6 and 11 are responsible for over 90% of genital warts and RRP cases, and 9 to 12% of low grade cervical dysplastic lesions.Identification of a viral agent such as HPV as a major cause of diseases impliesthat prophylactic vaccines or interventions against the viral agent should preventthe disease(s) it causes. Initial studies in animal models showed that inoculationwith species-specific papillomaviruses induced an immune response thatconferred protection against homologous virus challenge. However, nativepapillomaviruses are not good substrates for vaccine development as they cannot be grown easily in culture. Subsequent studies were initiated on the production of viral particles from expression of the structural proteins in heterologousexpression systems, such as yeast or baculovirus vectors. Results showed thatexpression of L1 alone led to the production of VLPs which morphologicallyresemble the authentic HPV virions but contain no viral DNA. These VLPs areproduced by self-assembly of the L1 protein when expressed in a heterologouscell substrate and are the basis for the vaccines considered in this document. Inanimal studies, VLPs were shown to protect against high dose experimentalinfection by homologous virus. HPV VLPs are highly immunogenic in mice orrabbits, and the resulting antibodies have been shown to be neutralizing and typerestricted when tested in a pseudovirion neutralization assay. Immunization withWHO/BS/06.2050 - FinPage 5 denatured particles does not result in the production of neutralizing antibodies, orprotect from experimental virus challenge, indicating that neutralizing epitopesare conformation dependent. Protection in animals has also been demonstratedthrough passive transfer of antibodies in serum.Neutralizing antibodies are probably the primary mediator of this protection. L1is not expressed in the basal keratinocytes in which infection is thought to bemaintained and regression of established lesions was not observed after VLPvaccination. Therefore, it seems unlikely that cell-mediated immunity (CMI) isinvolved as a direct effector mechanism of protection (5).The specific assays that have been developed to evaluate the immune responseinclude: VLP-based enzyme immunoassay (EIA), competitive immunoassay with labeled neutralizing monoclonal antibodies, hemagglutination inhibition (HAI),and in vitro neutralization.The development of these guidelines has been driven by the acquired experiencewith the two vaccines developed thus far. These vaccines are both made up ofrecombinant protein L1 VLPs and they contain adjuvant in order to stabilize theintegrity of the L1 VLPs and also to enhance immunogenicity. The productsdiffer in the types of HPV L1 proteins included as antigens, substrates used forproduction, adjuvant properties and in the final formulation. These two vaccinesare:1) A bivalent vaccine comprised of oncogenic HPV types 16 and 18 VLPsreassembled from L1 protein expressed and purified from insect cells infectedwith a recombinant baculovirus. This vaccine is formulated with a noveladjuvant, AS04, which contains aluminium hydroxide and monophosphoryllipid A (MPL); and2) A tetravalent vaccine comprised of the low risk HPV types 6 and 11 and theoncogenic HPV types 16 and 18. Type specific L1 proteins for this vaccineare expressed and purified from yeast cells containing L1 expression plasmids.The VLPs are adsorbed to an amorphous aluminium hydroxyphosphatesulfate-containing adjuvant.It is possible that a vaccine produced in mammalian cells may be developed in the future.Special considerationsThere are several special considerations that need to be addressed in themanufacturing, non-clinical and clinical development of these vaccine products.WHO/BS/06.2050 - FinPage 6VLPs are complex biological products and will need to be assessed at variouslevels.With respect to manufacturing and product quality the following items should beconsidered:1)The bivalent vaccine expressed from recombinant baculovirus in insect cells isthe first vaccine to be developed in this host expression system. Testing ofthis cell substrate may have some unique requirements;2) A novel adjuvant which has not previously been experienced on a global scaleis used in the formulation of the bivalent vaccine. The immunostimulant isMPL which is a detoxified form of lipid A derived from thelipopolysaccharide (LPS) isolated from bacterial cell walls of the Gramnegative bacterium Salmonella minnesota R595. While detoxified, MPL wasshown to retain the capacity of the natural LPS compound to act as animmunostimulant by potentiating cellular and humoral adaptive immuneresponses;3)L1 protein in its native form is not glycosylated. For the two current vaccinesglycosylation during production on a cell substrate is not an issue. HPV L1VLP vaccines produced in new or different cell substrates should be assessedfor glycosylation status;4)Disassembly and reassembly of the L1 capsomers may contribute topurification of the product and lead to more stable VLPs;5)Purified L1 VLP preparations will have to be characterized biochemically andimmunologically, to determine L1 concentration, purity and assembly state;and6)Current HPV vaccines are manufactured in single dose presentations withoutthe addition of preservative. In the future, the availability of multi-dosevaccine vials would facilitate the adoption of innovative vaccination strategiestargeting pre-adolescents and adolescents in developing countries. If thesevaccines do not contain preservative, the use of such vaccine vials should betime-restricted as is the case of reconstituted vaccines such as BacillusCalmette-Guérin (BCG) and measles-containing vaccines. If a preservativewere to be added, the effect on antigenicity and immunogenicity must beassessed and known not to have an negative impact as has been observed withthiomersal (6).With respect to the nonclinical studies it is critical that such studies demonstrateimmunogenicity and the production of neutralizing antibodies.With respect to clinical assessment of HPV VLP vaccines there are several critical considerations:WHO/BS/06.2050 - FinPage 71)Since 90% of HPV related cancers are cervical cancers, the efficacy of thevaccines developed so far has been studied in sexually active women;2)In order to obtain maximal benefit from these vaccines, the primary targetpopulation for immunization should consist of young adolescents prior toonset of sexual activity. Although the attack rate for HPV is high in the 5 to 10years following sexual debut, most women remain naïve to vaccine HPVtypes during this time, and few have been infected with all vaccine HPV types;3)Licensure of first generation vaccines requires a definitive demonstration ofprophylactic efficacy with respect to cervical intraepithelial neoplasm (CIN)2/3 and adenocarcinoma in situ (AIS) caused by vaccine HPV types;5)Persistent infection (e.g. detection of the DNA of the same virion incervicovaginal specimens collected on consecutive visits over a period of atleast 12 months) may be an appropriate endpoint for second generationvaccines, including those with additional HPV types. At the time of preparingthese Guidelines, however, there was no international consensus on adefinition for HPV persistence based on detection of HPV DNA by restrictedPCR; and6)Once licensed, long term effectiveness evaluation of these vaccines should beintegrated with current screening programs for cervical cancer.Part A. Guidelines on manufacturingA.1 DefinitionsA.1.1 International name and proper nameThe international name should be “Recombinant human papillomavirus virus-like particle vaccine” followed in parenthesis by the genotype specificity and the name of recombinant protein (e.g. genotype 16 and 18 L1 proteins). The proper nameshould be equivalent to the international name in the language of the country oforigin.The use of the international name should be limited to vaccines that satisfy thespecifications elaborated below.A.1.2 Descriptive definitionThe recombinant HPV VLP vaccine is a sterile liquid vaccine preparation whichcontains purified VLPs composed of the recombinant major capsid proteins ofone or more HPV genotypes (further referred to as "types"). The VLPs may beformulated with a suitable adjuvant. Such vaccines are for prophylactic use.A.1.3 International reference preparationsWHO/BS/06.2050 - FinPage 8International reference preparations based on recombinant HPV VLPs were notavailable when this Guidelines were prepared. However, reference reagents foruse in the laboratory evaluation of the biological effects following vaccineadministration to humans, such as antibody titers and viral DNA detection, areunder development for HPV types 16 and 18. Some information can be found inthe literature (7-10).A.1.4 TerminologyThe definitions given below apply to this document only.HPV L1 protein: The major structural protein of human papillomavirus, of which 360 molecules are found in the native virion associated in 72 pentamericcapsomers.L1 virus-like particle: A non-infectious, non-enveloped, icosahedral capsidparticle which does not contain viral DNA and which is composed of regulararrays of L1 pentameric capsomers.Parental yeast cell: Yeast host cell to be manipulated for the expression ofprotein(s) to give rise to a recombinant yeast production strain.Recombinant baculovirus master seed lot: A quantity of recombinant baculovirus of uniform composition derived from an original baculovirus construct, processed at one time and passaged for a documented number of times.Recombinant baculovirus working seed lot: A quantity of recombinantbaculovirus of uniform composition, derived from the master seed lot by a limited number of passages. The recombinant baculovirus virus working seed lot may be used to prepare inoculum intermediates or alternatively to initiate the productionof recombinant L1 proteins.Inoculum intermediate: A quantity of recombinant baculovirus of uniformcomposition, derived from the working seed lot. The inoculum intermediate has a defined shelf-life. It is intended to be used to initiate the production ofrecombinant L1 proteins.Cell bank: A collection of ampoules containing aliquots of a suspension of cellsfrom a single pool of cells of uniform composition, stored frozen under definedconditions (typically <−60 °C for yeast, and in liquid nitrogen for insect ormammalian cell lines).Master cell bank (MCB): A collection of containers containing aliquots of asuspension of cells from a single pool of cells of uniform composition, storedfrozen under defined conditions (typically <−60 °C for yeast, and in liquidWHO/BS/06.2050 - FinPage 9 nitrogen for insect or mammalian cell lines). The MCB is used to derive all working cell banks for the anticipated lifetime of the vaccine product.Working cell bank (WCB): A collection of containers containing aliquots of a suspension of cells from a single pool of cells of uniform composition, derived from the MCB, stored frozen under defined conditions (typically <−60 °C for yeast, and in liquid nitrogen for insect or mammalian cell lines). One or more aliquots of the WCB are used for routine production of the vaccine. Multiple WCBs are made and used during the lifetime of vaccine productProduction cell culture: A cell culture derived from one or more containers of the WCB used for the production of vaccines.End of production cells: A cell suspension containing the cells harvested at the end of culture/fermentation.Adventitious agents: Contaminating microorganisms of the virus, or cell substrate or materials used in their cultures, that may include bacteria, fungi, mycoplasmas, and endogenous and exogenous viruses that have been unintentionally introduced. Fermentation cell paste: A suspension of cells harvested at the end of the yeast fermentation stored frozen (<-60°C).Single antigen harvest: A cell-suspension containing the intended HPV antigens of one virus type harvested from cell cultures prepared from a single production runSingle harvest pool: A homogenous pool of multiple single harvests of the intended HPV antigens of one virus type, collected into a single vessel before clarification.Purified monovalent antigen bulk: A batch of purified antigen of the same HPV type. Different batches of purified monovalent antigen bulks may be pooled before collection into a single vessel.Adsorbed monovalent antigen bulk: A batch of purified monovalent antigen bulk adsorbed on an aluminium containing adjuvant. Different batches of adsorbed monovalent antigen bulks may be pooled before collection into a single vessel. Adjuvant: A vaccine adjuvant is a component that potentiates the immune response to an antigen and/or modulates it towards the desired immune responses.Final vaccine bulk: The formulated bulk present in the container from which the final containers are filled. The final bulk may be prepared from one or more adsorbed monovalent antigen bulks and may contain VLP antigens from one or multiple HPV virus types.WHO/BS/06.2050 - FinPage 10Filling lot (final vaccine lot): A collection of sealed final containers of vaccinethat is homogeneous with respect to the risk of contamination during the fillingprocess. A filling lot must therefore have been filled or prepared in one workingsession.A.2 General manufacturing recommendationsThe general manufacturing requirements contained in Good manufacturingpractices for biological products (11) should apply to the establishment ofmanufacturing facilities for recombinant HPV VLP vaccines, with the addition of the following:•Production steps involving manipulations of recombinant HPV L1 VLP types should be conducted at a biosafety level consistent with the recombinantproduction microorganism;•Quality control procedures should be in place to ensure segregation of different HPV L1 VLP types during bulk manufacturing steps.Sufficientcleaning validation and product changeover data should be available; and•The antigen manufacturing process should be validated to demonstrate production consistency. Typically, three consecutive lots per HPV type arerequired. However, if one or more HPV types use the same manufacturingprocess, validation of all processes with at least one type may be acceptable.The assessment of manufacturing consistency should include evaluation ofcritical quality parameters and their corresponding attributes. Examples ofprocess quality attributes are nucleic acid and host cell protein clearance orcumulated population doubling level and examples or process key operatingparameters is column loading. The process validation antigen batches shouldshow compliance with the pre-established antigen quality controlspecifications for the HPV antigen such as antigen identity and antigen purity(see section A.5).A.2.1 Characterization of the antigenCharacterization of HPV antigen is performed on lots produced during vaccinedevelopment, including the process validation batches.The protein composition should be established by techniques such as sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions or mass spectrometry. The bands should be identified by sensitivestaining techniques and where possible by specific antibodies or massspectrometry to confirm the presence of the expected products of the L1 protein.The identity of the protein should be established by peptide mapping and/orterminal amino acid sequence analysis.Page 11 Since it is known that conformational epitopes are essential for efficacy, it isessential that the morphological characteristics of the VLPs and degree ofaggregation should be determined. In addition, the protein, lipid, nucleic acid andcarbohydrate content should be measured when applicable. VLP characterizationmay be done by atomic force and transmission electron microscopy, dynamiclight scattering, epitope mapping and reactivity with neutralizing monoclonalantibodies.The level of residual host cell protein derived from insect cells and/or a novel cellsubstrate should meet acceptable safety in nonclinical and clinical studies (seeParts B and C).A.3 Control of source materialsA.3.1 Cell cultures for antigen productionThe use of any cell line should be based on a cell bank system. Only cells thathave been approved and registered with the national regulatory authority shouldbe used to produce HPV L1 protein. The national regulatory authority should beresponsible for approving the cell bank. Appropriate history of the cell bankshould be provided.A.3.1.1 Yeast cellsThe characteristics of the recombinant production strain (host cell in combinationwith the expression vector system) should be fully described and informationgiven on the absence of adventitious agents and on gene homogeneity for themaster and working cell banks. A full description of the biological characteristicsof the host cell and expression vectors should be given. The physiologicalmeasures used to promote and control the expression of the cloned gene in thehost cell should be described in detail. This should include genetic markers of the host cell, the construction, genetics and structure of the expression vector and theorigin and identification of the gene that is being cloned.The nucleotide sequence of the gene insert and of adjacent segments of the vectorand restriction-enzyme mapping of the vector containing the gene insert should beprovided as required by the national control authority.A.3.1.2 Insect cellsIf insect cells are used for production of recombinant HPV L1 VLP vaccines, theuse of insect cell substrate should be based on a cell bank system. The cellsubstrates and cell banks should conform with Requirements for use of animalcells as in vitro substrates for the production of biologicals (12,13), as appropriateto insect cells, and should be approved by the national regulatory authority.Page 12The maximum number of passages (or population doublings) allowable betweenthe MCB, the WCB and the production cells should be approved by the nationalregulatory authority. Additionally, the MCB or WCB cells should be propagatedto or beyond the maximum production level and be examined for tumorigenicityin an animal test system and for the presence of retroviruses and arthropod-borne viruses.The MCB is made in sufficient quantities and stored in a secure environment and is used as the source material to make manufacturers WCB. In normal practice aMCB is expanded by serial subculture up to a passage number (or populationdoubling, as appropriate) selected by the manufacturer and approved by thenational regulatory authority, at which point the cells are combined to give asingle pool distributed into ampoules and preserved cryogenically to form theWCB.The manufacturers working cell bank is used for the preparation of productioncell culture, and thus for production of HPV L1 antigen batches.A.3.1.3 Other Cell SubstratesIf other host cells are used, the cell substrates and cell banks should conform with Requirements for use of animal cells as in vitro substrates for the production ofbiologicals (12,13) where appropriate, and should be approved by the nationalregulatory authority.A.3.2 Cell culture mediumIf serum is used for the propagation of cells, it should be tested to demonstratefreedom from bacteria, fungi and mycoplasmas, according to the requirementsgiven in Part A, sections 5.2 (14) and 5.3 (15)of Requirements for biologicalsubstances no. 6 and from infectious viruses. Suitable tests for detecting viruses in bovine serum are given in Appendix 1 of Recommendations for production andcontrol of poliomyelitis vaccine (oral) (16).Validated molecular tests for bovine viruses may replace the cell culture tests ofbovine sera. As an additional monitor of quality, sera may be examined forfreedom from phage and endotoxin. Gamma-irradiation may be used to inactivate potential contaminant viruses.The acceptability of the source(s) of any components of bovine, porcine, sheep or goat origin used should be approved by the national regulatory authority. Thesecomponents should comply with current WHO guidelines in relation to animaltransmissible spongiform encephalopathies (17).。
WHO对HPV疫苗质量、安全性及有效性指导原则
欢迎阅读GUIDELINESTOASSURETHEQUALITY,SAFETYANDEFFICACYOFREC OMBINANTHUMANPAPILLOMA VIRUSVIRUS?LIKEPARTICLEV ACCIN ESHPV二、Specialconsiderationsection:在生产、非临床及临床中过程中的考虑因素：1、 VLP 1) 2)3) 4) L1VLP 。
5) 纯化后的L1VLP 要进行生化及免疫上的鉴定，并测定L1的浓度、纯度及组聚情况。
6) 如加入了防腐剂，应对其免疫性进行验证，并确认不会有负作用2、 非临床方面：关键就是要证明其免疫原性，并能否产生免疫中和抗体。
3、临床方面：（略）三、生产指导（PartA.Guidelinesonmanufacturing）3.1定义definitions3.1.1国际名称和专有名称3.1.2重组基因型3.1.33.2术语hichdoesnotcontainviralDNAandwhichiscomposedofregulararraysofL1pentamericcaps omers.Parentalyeastc ell:Yeasthostcelltobemanipulatedfortheexpressionofprotein(s)togiv erisetoarecombinantyeastproductionstrain.Inoculumintermediate:Aquantityofrecombinantbaculovirusofuniformcomposition, derivedfromtheworkingseedlot.Theinoculumintermediatehasadefinedshelf?life.Itisinten dedtobeusedtoinitiatetheproductionofrecombinantL1proteins.Cellbank:Acollectionofampoulescontainingaliquotsofasuspensionofcellsfromasin glepoolofcellsofuniformcomposition,storedfrozenunderdefinedconditions(typically<?6 0°Cforyeast,andinliquidnitrogenforinsectormammaliancelllines).Mastercellbank(MCB):Acollectionofcontainerscontainingaliquotsofasuspension ofcellsfromasinglepoolofcellsofuniformcomposition,storedfrozenunderdefinedconditioaterialsusedintheircultures,thatmayincludebacteria,fungi,mycoplasmas,andendogenous andexogenousvirusesthathavebeenunintentionallyintroduced.Fermentationcellpaste:Asuspensionofcellsharvestedattheendoftheyeastfermentat ionstoredfrozen(<?60°C).Singleantigenharvest:Acell?suspensioncontainingtheintendedHPVantigensofonevi rustypeharvestedfromcellculturespreparedfromasingleproductionrunSingleharvestpool:AhomogenouspoolofmultiplesingleharvestsoftheintendedHPVa ntigensofonevirustype,collectedintoasinglevesselbeforeclarification.Purifiedmonovalentantigenbulk:AbatchofpurifiedantigenofthesameHPVtype.Di fferentbatchesofpurifiedmonovalentantigenbulksmaybepooledbeforecollectionintoasin glevessel.3.3应分别生产，同时，还要有充分的清洁验证。
WHO对HPV疫苗质量安全性及有效性指导原则
WHO对HPV疫苗质量安全性及有效性指导原则世界卫生组织（WHO）对人乳头瘤病毒（Human Papillomavirus，简称HPV）疫苗的质量、安全性及有效性制定了一系列指导原则。
首先，WHO建议疫苗研发商在研发和生产过程中遵循国际标准和规范，确保疫苗的质量。
这包括对病毒株的选择、制备疫苗的工艺、生产设备和环境的控制等方面。
同时，疫苗研发商需要进行严格的质量控制，确保每一批疫苗都符合质量标准。
其次，WHO强调疫苗的安全性是至关重要的。
疫苗研发商需要在临床试验阶段进行全面的安全性评估，并依据国际规范进行监测和报告。
在疫苗上市后，监管机构和疫苗研发商需要持续监测疫苗的安全性，及时报告任何可能的不良反应，并采取必要的措施进行风险管理。
第三，WHO指导疫苗研发商进行有效性评估。
疫苗的有效性评估需要包括原始病毒株类型的选择、免疫原性和保护机制的研究、动物模型和临床试验等方面。
通过这些研究，可以评估疫苗的免疫原性和保护效果，从而确定疫苗是否能有效预防HPV感染和相关疾病。
此外，WHO还倡导疫苗研发商进行全球合作，共同推动HPV疫苗研发和应用，以降低世界各地HPV感染和相关疾病的发病率和死亡率。
为此，WHO提供技术支持和合作机会，促进疫苗研发商在全球范围内进行合作，共同提高HPV疫苗的质量、安全性和有效性。
最后，WHO强调对HPV疫苗的监管和监测工作的重要性。
国家和地区的监管机构需要确保疫苗研发商遵守相关法规和规范，对疫苗进行审查和批准，并对疫苗市场进行监管。
同时，监管机构需要与疫苗研发商、卫生机构和公众开展沟通和合作，共同推动HPV疫苗的安全性和有效性监测工作。
总结起来，世界卫生组织对HPV疫苗的质量、安全性及有效性制定了一系列指导原则，包括遵循国际标准和规范、进行严格的质量控制、全面评估疫苗的安全性和有效性、倡导全球合作和加强监管和监测工作等。
这些指导原则对于保证HPV疫苗的质量、安全性和有效性，进一步预防和控制HPV感染和相关疾病具有重要意义。
WHO对HPV疫苗质量、安全性及有效性指导原则
WHO对HPV疫苗质量、安全性及有效性指导原则GUIDELINES TO ASSURE THE QUALITY, SAFETY AND EFFICACY OF RECOMBINANT HUMAN PAPILLOMA VIRUS VIRUS¬LIKE PARTICLE V ACCINESHPV二、Special consideration section:在生产、非临床及临床中过程中的考虑因素：1、生产方面：VLP是复杂的生物产物，必须在不同水平下对其进行检测分析。
因而在其生产过程及质量控制上必须考虑以下几个因素：1)新的表达体系如杆状病毒（GSK），新的特殊要求。
但我们用的毕赤酵母，相对比较常见。
2)新佐剂（略）3)天然的L1蛋白是没有被糖基化修饰，目前的两种表达体系，在糖基化修饰上不存大问题,但要对糖基化及其位点进行分析。
4)L1衣壳蛋白亚单位的解聚与再聚，可能有利于纯化，并得到更稳定的VLP。
目前，我们的路线可能是不经解聚，直接纯化获得VLP。
个人感觉，到后期可以兵分两路，一路直接获得VLP，而另一路则将VLP解聚后，再进行纯化与重组。
5)纯化后的L1 VLP要进行生化及免疫上的鉴定，并测定L1的浓度、纯度及组聚情况。
6)如加入了防腐剂，应对其免疫性进行验证，并确认不会有负作用2、非临床方面：关键就是要证明其免疫原性，并能否产生免疫中和抗体。
3、临床方面：（略）三、生产指导（Part A. Guidelines on manufacturing）3.1定义definitions3.1.1国际名称和专有名称国际名称：重组人乳头瘤类病毒颗粒疫苗（基因型16 L1蛋白）3.1.2定义描述重组HPV VLP疫苗为无菌的液态疫苗，里面含纯化后由一种或多种HPV基因型重组的主要的衣壳蛋白，并与相应佐剂混合。
3.1.3国际标准品在此指导原则编写时，市场上暂无国际标准品提供。
但有相应试剂在实验室水平上，在进行注射后进行生物效价方面的评价如抗体滴度和病毒DNA检测。
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GUIDELINES TO ASSURE THE QUALITY, SAFETY AND EFFICACY OF RECOMBINANT HUMAN PAPILLOMA VIRUS VIRUS¬LIKE PARTICLE V ACCINESHPV二、Special consideration section:在生产、非临床及临床中过程中的考虑因素:1、生产方面:VLP就是复杂的生物产物,必须在不同水平下对其进行检测分析。
因而在其生产过程及质量控制上必须考虑以下几个因素:1)新的表达体系如杆状病毒(GSK),新的特殊要求。
但我们用的毕赤酵母,相对比较常见。
2)新佐剂(略)3)天然的L1蛋白就是没有被糖基化修饰,目前的两种表达体系,在糖基化修饰上不存大问题,但要对糖基化及其位点进行分析。
4)L1衣壳蛋白亚单位的解聚与再聚,可能有利于纯化,并得到更稳定的VLP。
目前,我们的路线可能就是不经解聚,直接纯化获得VLP。
个人感觉,到后期可以兵分两路,一路直接获得VLP,而另一路则将VLP解聚后,再进行纯化与重组。
5)纯化后的L1 VLP要进行生化及免疫上的鉴定,并测定L1的浓度、纯度及组聚情况。
6)如加入了防腐剂,应对其免疫性进行验证,并确认不会有负作用2、非临床方面:关键就就是要证明其免疫原性,并能否产生免疫中与抗体。
3、临床方面:(略)三、生产指导(Part A、Guidelines on manufacturing)3、1定义definitions3、1、1国际名称与专有名称国际名称:重组人乳头瘤类病毒颗粒疫苗(基因型16 L1蛋白)3、1、2定义描述重组HPV VLP疫苗为无菌的液态疫苗,里面含纯化后由一种或多种HPV基因型重组的主要的衣壳蛋白,并与相应佐剂混合。
3、1、3国际标准品在此指导原则编写时,市场上暂无国际标准品提供。
但有相应试剂在实验室水平上,在进行注射后进行生物效价方面的评价如抗体滴度与病毒DNA检测。
3、2术语TerminologyThe definitions given below apply to this document only、HPV L1 protein: The major structural protein of human papillomavirus, of which 360 molecules are found in the native virion associated in 72 pentameric capsomers、L1 virus¬like particle: A non¬infectious, non¬enveloped, icosahedral capsid particle which does not contain viral DNA and which is composed of regular arrays of L1 pentameric capsomers、Parental yeast c ell: Yeast host cell to be manipulated for theexpression of protein(s) to give rise to a recombinant yeast production strain、Inoculum intermediate: A quantity of recombinant baculovirus of uniform composition, derived from the working seed lot、The inoculum intermediate has a defined shelf¬life、It is intended to be used to initiate the production of recombinant L1 proteins、Cell bank: A collection of ampoules containing aliquots of a suspension of cells from a single pool of cells of uniform composition, stored frozen under defined conditions (typically <−60 °C for yeast, and in liquid nitrogen for insect or mammalian cell lines)、Master cell bank (MCB): A collection of containers containing aliquots of a suspension of cells from a single pool of cells of uniform composition, stored frozen under defined conditions (typically <−60 °C for yeast, and in liquid nitrogen for insect or mammalian cell lines)、The MCB is used to derive all working cell banks for the anticipated lifetime of the vaccine product、Working cell bank (WCB): A collection of containers containing aliquots of a suspension of cells from a single pool of cells of uniform composition, derived from the MCB, stored frozen under defined conditions (typically <−60 °C for yeast, and in liquid nitrogen for insect or mammalian cell lines)、One or more aliquots of the WCB are used for routine production of the vaccine、Multiple WCBsare made and used during the lifetime of vaccine productProduction cell culture:A cell culture derived from one or more containers of the WCB used for the production of vaccines、End of production cells: A cell suspension containing the cells harvested at the end of culture/fermentation、Adventitious agents: Contaminating microorganisms of the virus, or cell substrate or materials used in their cultures, that may include bacteria, fungi, mycoplasmas, and endogenous and exogenous viruses that have been unintentionally introduced、Fermentation cell paste: A suspension of cells harvested at the end of the yeast fermentation stored frozen (<¬60°C)、Single antigen harvest: A cell¬suspension containing the intended HPV antigens of one virus type harvested from cell cultures prepared from a single production runSingle harvest pool: A homogenous pool of multiple single harvests of the intended HPV antigens of one virus type, collected into a single vessel before clarification、Purified monovalent antigen bulk: A batch of purified antigen of the same HPV type、Different batches of purified monovalent antigen bulks may be pooled before collection into a single vessel、Adsorbed monovalent antigen bulk: A batch of purified monovalent antigen bulk adsorbed on an aluminium containingadjuvant、Different batches of adsorbed monovalent antigen bulks may be pooled before collection into a single vessel、Adjuvant: A vaccine adjuvant is a component that potentiates the immune response to an antigen and/or modulates it towards the desired immune responses、Final vaccine bulk:The formulated bulk present in the container from which the final containers are filled、The final bulk may be prepared from one or more adsorbed monovalent antigen bulks and may contain VLP antigens from one or multiple HPV virus types、Filling lot (final vaccine lot): A collection of sealed final containers of vaccine that is homogeneous with respect to the risk of contamination during the filling process、A filling lot must therefore have been filled or prepared in one working session、3、3生产建议生产必须符合GMP要求,生物安全上要求无菌。