Case Studies of Microbial Contamination in Biologic Product Manufacturing

Transcription

1 Microbiology» Case Studies of Microbial Cotamiatio i Biologic Product Maufacturig Kalavati Suvara, Ph.D, Aastasia Lolas, M.S. Patricia Hughes, Ph.D. & Richard L Friedma, M.S. Biotechology Maufacturig Team, Divisio of Maufacturig ad Product Quality, Office of Compliace, Ceter for Drug Evaluatio ad Research, Food ad Drug Admiistratio [email protected]. Abstract The maufacture of biologic products is a complex process ad requires the use of livig cells. These processes ad products are proe to cotamiatio by advetitious agets such as bacteria, fugi ad viruses. Microbial cotamiatios have a huge impact o biologic product maufacture as they itroduce product variability ad ca cause loss of potecy due to degradatio or modificatio of product by microbial ezymes, chages i impurity profiles, ad a icrease i the levels of bacterial edotoxis. I additio, the ivestigatios of microbial cotamiatios ca result i legthy shutdow periods ad delays i maufacturig operatios that i tur, may sometimes result i shortages of essetial drug products. Strict microbial productio cotrols are essetial to esure the maufacture of a drug product with cosistet quality. This article discusses elemets of a microbial cotrol strategy, recet cases of microbial cotamiatio i specified biologic products, the eed to perform risk assessmets o a periodic basis, ad additioal areas of improvemet i the maagemet of risks. Itroductio Biologic products are maufactured usig livig cells such as bacteria, yeast, ad mammalia cells. These iclude specified biologics such as moocloal atibodies ad therapeutic recombiat DNA-derived products licesed uder Sectio 351 of the Public Health Service Act [1] ad curretly regulated by the Ceter of Drug Evaluatio ad Research (CDER). These biological products are also regulated as drugs uder the Federal Food, Drug, ad Cosmetic Act [2]. The upstream process i the maufacture of moocloal atibodies ad therapeutic recombiat proteis typically ivolves cell expasio, cell culture, ad recovery steps. The dowstream process ivolves multiple purificatio steps. The purified protei is ultrafiltered/diafiltered with formulatio buffer to provide a formulated bulk drug substace. The formulated bulk drug substace is sterile-filtered ad filled to provide a fial drug product. Because of the cosequeces of microbial cotamiatio o product safety ad quality, there is cotiued iterest i uderstadig the root causes of microbial cotamiatio ad cotrollig these risks i biologic product maufacture. This article discusses some of the bacterial cotamiatio cases reported to the Agecy or idetified durig pre-licese/pre-approval ispectios of biologic drug substace maufacturers i the past two years. The cases highlight areas for 50 Jauary/February 2011

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3 Microbiology» improvemet i risk maagemet ad the eed for developig a robust microbial cotrol strategy for biologic products. Sources of Microbial Cotamiatio Microorgaisms are ubiquitous i ature. Microorgaisms ca adapt ad survive uder a variety of coditios ad ca pose a sigificat risk to biologic products. A uderstadig of the microbial etry poits ad implemetatio of measures to prevet microbial cotamiatio is critical for maufacture of safe, pure ad potet biologic products. As show i Figure 1, microorgaisms ca gai etry ito a productio process stream from several sources: the facility, equipmet, process operatios, raw materials, colum resis, filter membraes, water, process gases, ad persoel. All sources of microbial cotamiatio should be cosidered whe developig a microbial cotrol strategy ad performig a ivestigatio for a microbial cotamiatio deviatio. address the quality cocers origiatig from cell substrates used for maufacture of these products. These documets iclude (a) Poits to Cosider i the Maufacture ad Testig of Moocloal Atibody Products for Huma Use published i February 1997, (b) Guidace o Viral Safety Evaluatio of Biotechology Products Derived From Cell Lies of Huma or Aimal Origi (Q5A), (c) Guidace o Quality of Biotechological/Biological Products: Derivatio ad Characterizatio of Cell Substrates Used for Productio of Biotechological/Biological Products (Q5D), ad (d) Guidace o Specificatios: Test Procedures ad Acceptace Criteria for Biotechological/Biological Products (Q6B) [6,7,8,9]. The Q6B guidace states that cotamiats should be strictly avoided ad/or suitably cotrolled with appropriate i-process acceptace criteria or actio limits for the drug substace or drug product to meet specificatios. The 1994 FDA Guidace for Idustry o the Submissio Documetatio for Sterilizatio Process Validatio i Applicatios for Huma ad Veteriary Drug Products provides guidace o sterilizatio process validatio for fial drug product [10]. The 2004 FDA Guidace for Idustry o Sterile Drug Products Produced by Aseptic Processig Curret Good Maufacturig Practice provides guidace o persoel qualificatios, clea room desig, process desig, ad aseptic processig of fial drug products [11]. These regulatios ad guidace documets provide the backboe for the developmet of a appropriate microbial cotamiatio cotrol strategy. Elemets of a Microbial Cotrol Strategy Figure 1: Sources of microbial cotamiatio. Regulatio ad Guidace The miimum curret good maufacturig practice (CGMP) requiremets for preparatio of fiished huma drug products are described i 21CFR 211 [3]. These iclude the use of suitable protective apparel (21CFR ), appropriate facility desig ad placemet of equipmet (21CFR ), equipmet cleaig, sterilizatio, ad maiteace (21CFR ), ad productio ad process cotrols (21CFR ). All these prevetive measures ad precautios are implemeted to protect product ad prevet cotamiatio. Therapeutic recombiat products ad moocloal atibodies are also subject to applicable regulatio i 21CFR parts [4]. The guidace o CGMP for active pharmaceutical igrediets, Q7A, provides geeral CGMP guidace for biologic drug substace maufactured by cell culture or fermetatio uder sectio XVIII [5]. Additioal guidace documets cover prevetio or cotrol of advetitious agets i cell-derived biologic products ad A microbial cotrol strategy should be developed oce a comprehesive risk assessmet has bee performed for all possible microbial etry poits ito the maufacturig process. This requires a good uderstadig of the maufacturig process ad product attributes. I geeral, the desig of the facilities should allow for proper operatios ad prevetio of cotamiatio. The flow of persoel, material ad waste should be from clea to dirty areas ad critical upstream ope operatios liable to microbial cotamiatio should be performed i desigated biosafety hoods or areas with ISO 5 classificatio. Depedig o the risks to the process, areas should be appropriately segregated. Segregatio of pre-viral ad post-viral clearace steps i processes usig mammalia host cells is importat to prevet cross-cotamiatio of process itermediates ad the facility. Segregatio of areas, appropriate chageover procedures, ad other procedural cotrols should be i place to prevet crosscotamiatio i a multi-product facility. Evirometal moitorig of maufacturig areas should be performed routiely at appropriate itervals. Process gases ad water should be tested ad moitored to esure adequate microbial cotrol. The desig of equipmet (sigleuse disposable versus multi-use), validated cleaig ad sterilizatio processes alog with a comprehesive prevetative maiteace pla are critical compoets of the microbial cotrol strategy. Microbial cotrol for the lifetime use of membraes ad resis should be demostrated. I additio, it is critical to idetify ad establish processig steps that decrease bioburde ad bacterial edotoxi levels as the process itermediates are processed through sequetial purificatio steps. Bioburde reducig filters should be used at 52 Jauary/February 2011

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5 Microbiology» critical steps i the process. This is critical for buffer solutios ad i process itermediates coducive to microbial growth. Miimizig the umber of ope operatios reduces the risk to product from exteral (persoel ad evirometal) microbial cotamiatio sources. Biologic products are usually rich i carbo sources that favor microbial growth. Hold coditios (time, temperature) for a process should be validated to cotrol ad prevet potetial microbial growth. Bioburde ad edotoxi alert ad actio limits should be set for process steps based o process capability. Raw materials should be screeed for microbial quality ad should be hadled ad stored i a maer to prevet cotamiatio ad crosscotamiatio. Persoel are importat cotributors to microbial cotamiatios. Appropriate gowig should be implemeted to prevet cotamiatio. All persoel performig ope operatios should be traied adequately ad evaluated periodically i such operatios. Case Studies I the last two years, several cotamiatio evets were reported to the Agecy. They icluded viral or bacterial cotamiatio of upstream cell culture or fermetatio processes. Viral cotamiatio evets were extesively covered i the recet 2010 PDA/FDA Advetitious Viruses i Biologics: Detectio ad Mitigatio Strategies Workshop. Oly Itroducig AccuPRO-ID Discover the Power AccuPRO-ID is the ideal choice for routie evirometal moitorig ad primary screeig of bacterial orgaisms. Backed by our validated library, AccuPRO-ID outperforms the VITEK 2 Compact by 2X i accuracy. AccuPRO-ID vs. VITEK 2 Compact ASM 2010 Poster I-2664 = 60 ukow evirometal isolates (46 differet species) bacterial cotamiatios are discussed i this article. Oe case ivolved cotamiatio of a fermetor used i the maufacture of a protei product secreted by a bacterial host. The cotamiat was idetified as Bacillus cereus (a Gram positive spore formig rod). A secod case ivolved the cotamiatio of a fermetor used i the maufacture of a recombiat protei by Paeibacillus curdlaolyticus (a Gram variable spore-formig rod). A systematic approach was used durig the ivestigatios to idetify the root cause of the cotamiatio ad icluded several media simulatios to aid i idetifyig the poit of etry ito the fermetor. I additio, the ivestigatios ivolved the maufacture of egieerig batches. After a legthy ivestigatio i both case studies, problems with the samplig devices, additio valves, icorrectly fitted compoets, missig O-rigs, icorrect istallatio ad deformatio of a air filter after sterilizatio, ad/or iadequate slope of a codesate lie were idetified. Immediate corrective actios icluded the replacemet of valve diaphragms i fermetor additio ports, replacemet of a membrae valve i the samplig device, ad replacemet of O-rigs o the measurig probes. Ehacemets were also made to the sterilizatio processes of fermetor ad associated trasfer lies. A prevetative maiteace pla was developed for all fermetor valves. All valves were tagged usig a detailed checklist to esure correct istallatio. All SOPs were updated ad employees were traied o the revised versios. The ivestigatios ad corrective actios addressed all possible causes of cotamiatio as a uequivocal root cause could ot be assiged. I most cases, it is very difficult to idetify a defiitive assigable cause. It is highly recommeded that a systematic approach be followed to determie the root cause. Media simulatios help i demostratig that sterility of the fermetor is ot compromised. Recet microbial cotamiatio evets at several maufacturig facilities poit to breaches i the sterile boudary caused by damaged vet filters, damaged O-rigs, diaphragms, ad elastomers, ad improperly sloped Higher accuracy at lower cost to customers curretly usig ay pheotypic method Utilizes our validated, cgmp libraries, based o testig hudreds of thousads of idustry-relevat orgaisms Samples that do ot produce a result usig MALDI-TOF will automatically be sequeced for a ID at o additioal charge Try us out for free! Simply submit up to 5 bacterial samples ad write the Redemptio Code: APR-AD i the Commets colum o the Idetificatio Request Form. To access the form, please visit us at ad dowload AccuPRO-ID IRF or call Image: Copyright Deis Kukel Microscopy, Ic,. codesate lies. Whe bacterial hosts are used, microscopic examiatios of the fermetatio culture for cotamiatio is difficult. A culture purity test should be perfomed usig appropriate media ad culture coditios. It is crucial to have a comprehesive prevetative maiteace pla for fermetor ad tak agitators, probes, gaskets, O-rigs, valves, ad filters. The desig of pipig ad valves should prevet steam codesate from collectig ad leadig to cotamiatio by back-flow. After periods of shutdow or maiteace, it is importat to perform media simulatios o sterile equipmet that has remaied idle for a period of time. Procedural details o assembly ad set-up of fermetors/ bioreactors should be clear ad very detailed. Traiig i this area ca reduce iadvertet 54 Jauary/February 2011

6 «Microbiology leaks ad cotamiatio of the systems. Cotiuous assessmets of chage cotrol, work orders, ad other process improvemets should be coducted to esure that the microbial cotrol strategy is ot impacted. Of ote i both cases, the cotamiatig microorgaism was a facultative aaerobic Gram positive spore-formig rod. Risk mitigatio strategies based o microbial evirometal flora should be cosidered. The areas for improvemet idetified i the case studies were i prevetative maiteace plas for all fermetor valves icludig valves o samplig devices ad i the documetatio for correct assembly of compoets. i the ivestigatios. A hazard aalysis ad critical cotrol poit assessmet for bioburde cotrol throughout the maufacturig process is useful for the desig of a microbial cotrol strategy ad the performace of a systematic ivestigatio. I additio, failure data should be tracked to gai a better uderstadig of root causes. The iformatio should be used to cotiuously evaluate risks ad implemet process ad/or equipmet improvemets to mitigate ad prevet microbial cotamiatios. Two cases of microbial cotamiatio of the dowstream process were idetified durig pre-approval/pre-licese ispectios of drug substace maufacturig facilities. Bioburde deviatios were observed i several batches at the ultra-filtratio/diafiltratio (UF/DF) step. The cotamiats idetified were Sphigomoas species, Steotrophomoas maltophilia, Ralstoia pickettii, ad Staphylococcus species suggestig probable water ad huma sources of cotamiatio. Presece of repeated high bioburde couts i several batches suggested developmet of biofilm ad iadequate cotamiatio cotrol procedures for the UF/DF steps. After extesive ivestigatios, several corrective actios were implemeted i terms of cleaig, storage ad re-use of UF/ DF systems, sterilizatio/saitizatio of buffer taks, assessmet of the water for ijectio (WFI) system ad trasfer lies, itroductio of i-process bioburde reducig filters (i cases where there were o filters before the UF/DF steps), validatio of hold times ad storage coditios of process itermediates ad revisios to bioburde limits based o process capability. Demostratio of microbial cotrol over the lifetime use of membraes ad validatio of i-process hold times are essetial for esurig the cosistet quality of biologic products. All WFI pipig locatios with stagat water should be assessed ad elimiated. Microbial tred reports for water systems should be reviewed regularly. The ivestigatios of microbial cotamiatios are challegig due to the ubiquitous ature of the microorgaisms, multiple poits of microbial etry, growth promotig properties of biological process streams, limitatios of samplig ad detectio methods, ad the time ad resources ivolved i performig complex ivestigatios. All microbial etry poits should be systematically evaluated. For fermetor cotamiatios, seed fermetors ad associated additios ad trasfer lies should be icluded Accelerate ad automate your microbial testig. The Growth Direct TM System Rapid Automated Compedial Test No-Destructive Replaces huma eye with digital imagig to deliver rapid microbial eumeratio: Automated icubatio, eumeratio, reportig & LIMS data trasfer saves labor & improves compliace Applicatios & throughput cover your routie QC testig eeds Streamlied validatio compatible with USP methods ad regulatory guidelies No-destructive test allows faster ivestigatios Save time. Save moey. Icrease productivity. Rapid Micro Biosystems, Ic. Bedford, MA Phoe: Jauary/February

7 Microbiology» Coclusios Microbial cotamiatio is a risk to biologic product quality ad safety. The cost of iadequate microbial cotrol i biologic product maufacture is eormous as facilities or bioreactor productio trais may have to be shut dow for legthy periods of time i order to coduct ivestigatios ad idetify the root cause to prevet reoccurrece. The recet cases of bacterial cotamiatio of biologic products suggest that prevetative maiteace plas for fermetor ad associated valves, types of materials used for diaphragms ad O-rigs, ad uderstadig of microbial cotrol at certai process steps eed further attetio. Cotamiatio cotrol requires a uderstadig of the microbial etry poits ad risks to the process as well as the microbial growth potetial of the product, media ad buffer solutios. Microbial cotamiatio cotrol requires appropriate desig of facility ad equipmet, validated cleaig ad sterilizatio cycles for equipmet, detailed ad robust prevetative maiteace plas for equipmet, measures to reduce bioburde ad bacterial edotoxis at appropriate steps i the process, ad routie moitorig of these process steps for bioburde ad edotoxi with defied alert ad actio limits. A cotamiatio remediatio pla should be established. Such a pla is beeficial for meetig CGMP ad has the advatage of reducig facility dowtime. Ivestigatios should be comprehesive ad iclude assessmet of all microbial etry poits. Corrective actios should address all possible idetified causes i the absece of a kow assigable root cause. The iformatio gathered durig these ivestigatios should feed ito the overall risk maagemet pla. The quality risk maagemet pla should be itegrated ito the quality system ad allow for cotiuous improvemet. Refereces 1. Public Health Service Act, Biological Products; as ameded 2. Federal Food Drug ad Cosmetic Act; as ameded. 3. FDA, Curret Good Maufacturig Practices for Fiished Pharmaceuticals, 21 CFR part FDA, Biologics, 21 CFR parts U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Guidace for Idustry: Q7A Good Maufacturig Practice Guidace for Active Pharmaceutical Igrediets. Rockville, MD; U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Cetre for Biologics Evaluatio ad Research. Poits to Cosider i the Maufacture ad Testig of Moocloal Atibody Products for Huma Use. February U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Guidace for Idustry: Q5A Viral Safety Evaluatio of Biotechology Products Derived From Cell Lies of Huma or Aimal Origi. Rockville, MD; U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Guidace for Idustry: Q5D Guidace o Quality of Biotechological/Biological Products: Derivatio ad Characterizatio of Cell Substrates Used for Productio of Biotechological/Biological Products. Rockville, MD; Federal Register Vol. 63, No. 182, U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Guidace for Idustry: Q6B Specificatios: Test Procedures ad Acceptace Criteria for Biotechological/Biological Products, FDA, U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Guidace for Idustry for the Submissio Documetatio for Sterilizatio Process Validatio i Applicatios for Huma ad Veteriary Drug Products. Rockville, MD; U.S. Departmet of Health ad Huma Services, Food ad Drug Admiistratio. Guidace for Idustry: Sterile Drug Products Produced by Aseptic Processig Curret Good Maufacturig Practice. Rockville, MD; Biography Kalavati Suvara, Ph.D. is a Microbiologist with the Biotech Maufacturig Team i the Divisio of Maufacturig ad Product Quality i the Office of Compliace, CDER, FDA. She has over ie years of experiece as a microbiology reviewer at the FDA. Kalavati holds a Ph.D. i Biological Scieces from Norther Illiois Uiversity. Prior to joiig the Agecy, she worked i a academic ad pharmaceutical settig. Aastasia G. Lolas is a Microbiologist with the Biotech Maufacturig Team i the Divisio of Maufacturig ad Product Quality i the Office of Compliace, CDER, FDA. She has over 5 years of experiece as a microbiology reviewer of drug applicatios at the FDA. Aastasia holds a B.S. i Biology from Virgiia Polytechic Istitute ad State Uiversity ad a M.S. i Food Sciece from the Uiversity of Illiois at Urbaa- Champaig. Patricia F. Hughes, Ph.D. is the Team Leader i the Biotech Maufacturig Team i the Divisio of Maufacturig ad Product Quality i the Office of Compliace i CDER, FDA. She has over twety years experiece i the Pharmaceutical/Biotech idustry i fermetatio & cell culture process developmet ad maufacturig. I additio, she has over twelve years of experiece as a microbiology reviewer at the FDA, i CDER ad CBER. Patricia holds a Ph.D. i Microbiology from Georgetow Uiversity. Richard Friedma is the Director of the Divisio of Maufacturig & Product Quality i the Ceter for Drug Evaluatio ad Research (CDER), Office of Compliace. I this positio, he directs the iterpretatio ad developmet of CGMP policy, review of ispectioal recommedatios ad determiatio of maufacturig site acceptability. He has bee employed by FDA sice 1990, icludig prior positios as New Jersey District Drug Specialist, CDER Seior Compliace Officer ad Team Leader of Guidace ad Policy. Mr. Friedma has authored several publicatios o topics icludig sterile drugs ad quality maagemet systems, ad was awarded The George M. Sykes Award by the Pareteral Society for outstadig joural paper for the year Mr. Friedma is also a adjuct faculty member of Temple Uiversity School of Pharmacy i their QA/RA graduate program. Prior to joiig FDA, Mr. Friedma worked i the toxicology research divisio of a iovator pharmaceutical compay. Mr. Friedma received his B.S. i Biology with hoors from Motclair State Uiversity i 1989 ad his M.S. i Microbiology from Georgetow Uiversity School of Medicie i May, Jauary/February 2011

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