Managing Your Environmental Isolates

Transcription

1 Managing Your Environmental Isolates

2 The Controversy Good practice includes the periodic challenge of prepared media with low levels of organisms. This includes USP indicator organisms as well as normal flora. -U.S. Food and Drug Administration: Guide to Inspections of Microbiological Pharmaceutical Quality Control Laboratories (1993)

3 After Detection, Identification is Key It is important to know: What is in the cleanroom environment The effectiveness of cleaning and sanitization procedures The source of the contaminant

4 Mass Spectrometry Biochemical methods Characterization Outsource Services Molecular Methods Identification Methods Genotypic Phenotypic Trending Tracking Data Mgmt PCR Involves sequencing regions of organism s ribosomal RNA genes Strain typing Can differentiate between many more stains than biochemical methods MALDI-TOF Uses protein patterns Rapid and accurate Can identify many more species than biochemical methods Biolog Many others Results can help determine nutrient requirements of the organism. Gram stain Colony morphology Inexpensive, quick

5 Sources of Contamination Personnel Utilities Raw Materials Facility Common culprits: Personnel Air Water Processes Processes Equipment Case Studies of Microbial Contamination in Biologic Product Manufacturing Suvama, et al., American Pharmaceutical Review, 2011.

6 Tracking and Trending Effective tracking and trending analysis is essential An early warning signal for system failures Provides clues to contamination source

7 Is it Objectionable? 21 CFR (d)(6) Each lot of a component, drug product container, or closure with potential for microbiological contamination that is objectionable in view of its intended use shall be subjected to microbiological tests before use. 21 CFR (a) Appropriate written procedures, designed to prevent objectionable microorganisms in drug products not required to be sterile, shall be established and followed. 21 CFR (b) There shall be appropriate laboratory testing, as necessary, of each batch of drug product required to be free of objectionable microorganisms. What is an Objectionable Organism? Sutton, Scott, Ph.D., American Pharmaceutical Review, October 2012.

8 Is it Objectionable? Ultimately, the laboratory must determine if the organism is objectionable Sterile vs. non-sterile products Intended use Intended recipient Intended route of administration

9 Managing Risk with Testing Disinfectant Challenge Test Growth Promotion Test Antimicrobial Effectiveness Test

10 Disinfectant Challenge Testing the most frequently isolated microorganisms from an environmental monitoring program may be periodically subjected to use-dilution testing with the agents used in the disinfection program to confirm their susceptibility, as there are real differences among different species in resistance to the lethal effects of different sanitizers. USP 38 <1072> Disinfectants and Antiseptics, 2015

11 Disinfectant Challenge Testing Routinely used disinfectants should be effective against the normal vegetative flora recovered from the facility A sound disinfectant program also includes a sporicidal agent. Guidance for Industry Sterile Drug Products Produced by Aseptic Processing, 2004

12 FDA Warning Letter Your response states that a supplemental disinfectant efficacy, using mold spores of in-house isolates on various surfaces will be completed by We suggest that this proposed study by conducted as soon as possible. FDA Warning Letter, July 12, 2012

13 FDA Observation The firm was unable to provide scientific rationale for use of the selected organism used in the Disinfectant Efficacy Study. These organisms were not representative of organisms isolated from the facility nor were they representative of the USP guidelines. FDA 483 Observation, May 25, 2011

14 Growth Promotion Testing The media used in the following tests should be capable of growing the organism: Environmental monitoring Media fills Sterility Test

15 Growth Promotion Testing The growth promotion test is a procedure used to demonstrate the media in the microbiological environmental monitoring program or in media-fill runs, are capable of supporting growth of indicator microorganism and of environmental isolates from samples obtained through the monitoring program or their corresponding ATCC strains. USP 38 <1116> Aseptic Processing Environments, 2015

16 Growth Promotion Testing In addition to standard bacterial and fungal species used for growth promotion and sterility test procedures, validation studies should demonstrate that bacterial or fungal species found in the production environment (such as environmental isolates that may have become resistant to disinfection procedures and production strains) are detectable by the method used. World Health Organization: Environmental Monitoring of Clean Rooms in Vaccine Manufacturing Facilities (2012)

17 Growth Promotion Testing Your firm does not perform challenge testing to the sterility media with environmental isolates from the environmental monitoring program. FDA Warning Letter, October 29, 2010

18 Antimicrobial Effectiveness Testing The standard battery of challenge organisms need not prevent the inclusion of other species of microorganisms if deemed useful to measure the biological activity of the preservative system for a specific product. USP <51> Antimicrobial Effectiveness Testing, 2015 Product Product

19 Antimicrobial Effectiveness Testing Products of any category whose formulas establish more extreme environmental conditions (ph, water activity, specific formula conditions) may warrant addition of isolates capable of survival if not growth under such extreme conditions allowing measure of preservative efficacy versus intrinsic hostility. Antimicrobial Effectiveness Testing: Culture Selection and Inoculum Preparation, Phil Geiss, 2013

20 Why Not Just Use Reference Cultures? Your isolates can be more resistant to a disinfectant The isolates may need special conditions to grow Isolates can be harder to detect in your product

21 Wild Strains versus Domesticated Strains Wild microorganisms are able to modify their properties according to their nutrient supply (for example, phase variation in bacteria) and, in particular, are able to adapt to limited nutrients. Multicellular microorganisms: laboratory versus nature, Zdena Palková, EMBO Reports, 2004 There is a strong argument that environmental isolates are the best challenge to media and for validation studies like sterility test validation. They are the most sensitive microorganisms, having become recently exposed to disinfectants, particular soils etc. The Use of Environmental Isolates, Tim Sandle, Pharmaceutical Microbiology blog, January 10, 2010

22 Preserving Isolate Characteristics Professional Preservation Services Long-Term Preservation Methods Short-Term Preservation Methods

23 Short-Term Preservation Methods Preservation Method Pros Cons Sub-culturing/ Refrigeration + Low tech - Highest risk of mutation - Risk of contamination - Must remain within 5 passages per USP Sub-zero Freezing (-20 C) + Viability can be maintained for 1-2 yrs. - Ice crystals and electrolyte fluctuations can damage organism - Risk of contamination - Freezer cost and maintenance

24 Long-Term Preservation Methods Preservation Method Ultra-low Freezing Cryogenic Freezing Pros + Reduces probability of mutation + Longer survival rate Cons - Labor intensive - Costly - Must closely monitor temperature - Vulnerable to power outages and failures Lyophilization + Reduced risk of intracellular ice crystallization; halts all enzymatic and non-enzymatic reactions + Easy storage - Requires specialized equipment - Labor intensive - Experimentation required; strains react differently to process

25 Professional Preservation Services Turn-key processes for isolate management Isolate identification Isolate manufacturing Isolate storage Characterizatio n and seed preparation Customized format production Shipping, distribution and storage

26 Step 1: Characterization and Preproduction Strain characterization Determine final isolate specifications for testing Media and conditions should be mimicked Packaging needs Distribution requirements Establish a project timeline

27 Step 2: Production Seed stock is established 3-phase quality control checks Before preservation After preservation After packaging Seed stock for storage and future use Stock for immediate production

28 Phenotypic & Genotypic ID Service Charles River AccuGENX-ID Complete DNA sequencing report Sample compared to a validated library of over 8,000 bacterial strains Critical for root-cause investigations

29 Step 3: Shipping and Storage Isolate is packaged, shipped and distributed Isolates are maintained for future use

30 Cost Analysis for Outsourcing Costs of Manual Prep Labor (hrs x wages) - Handling isolates - Quantitating samples - Identification for mutation - QC for contamination - Record keeping for audits Freezer 3 rd party identification Laboratory supplies Higher Cost Vs. Costs of Outsourcing Project price Identification (included) Maintenance & storage (included) Lower Cost

31 Considerations for Outsourcing Accreditation Experience Test-ready format options Quality reputation Finished goods storage Technical support and service Distribution options Product warranty

32 Final Thoughts All environmental isolates pose risk in aseptic manufacturing Current regulations require the use of isolates in testing Managing isolates can be difficult and expensive, but there are accredited providers to help

33 Questions Contact Charles River Complete our easy online isolate request form at criver.com/qcsolutions Include isolate name and species Include your desired isolate format Technical Assistance: Phone: Website:

34 Thank you!