Characterization of Microbial Isolates from the Manufacturing Process Stream

Characterization of Microbial Isolates from the Manufacturing Process Stream

2011 PMF Conference on GMP in Non-Sterile ProductionMarch 17-18

Iselin, New Jersey

2


• What is the “Manufacturing Process Stream”?

• What is “Characterization”?

• Why Characterize Your Microbial Isolates?

3


“So far, no regulatory authority has set formal microbial-control standards for the manufacture of nonsterile dosage forms. FDA’s policy is that individual firms must establish the environmental controls necessary for these products… Even after USP publishes its informational chapter titled “Microbiological Control Programs for Manufacturing Non-Sterile Pharmaceutical Drug Product,” individual companies will be free to strengthen their own monitoring regimes or not, according to their own judgments. The debate about the need for enhanced control and monitoring of nonsterile products likely will continue, but both sides seem to agree that decisions ultimately should be based on an analysis of products’ risk to the patient.”

“Manufacturers Recalibrate Microbial Control for Nonsterile Drugs”

Erik GrebPharmaceutical Technology, February 16, 2011

Why are we here?

4


What is the “Manufacturing Process Stream”?

5


The Manufacturing Process Stream isn’t just the “Plumbing”

6


Raw Materials

and Components

Manufacture

Final Product

Facility

Utilities

Personnel

Equipment

Testing

Testing

Storage

Testing

Stability

7


“Case Studies of Microbial Contamination in Biologic Product Manufacturing”Kalavati Suvarna, Ph.D, Anastasia Lolas, M.S., Patricia Hughes, Ph.D. and Richard L

Friedman, M.S.American Pharmaceutical Review, January/February 2011

8


•Inherent bioburden•Susceptibility to contamination•Ability to support microbial growth

Testing

•Containment•Temperature•Humidity

Storage

Raw Materials and Components

9


•Environment (HVAC)•Surfaces•Cleaning (Disinfectants)

Facility

•Water (Potable, DI, etc.)•GasesUtilities

•Gowning•FlowPersonnel

•Tanks•“Plumbing”•Cleaning

Equipment

• In-process monitoringTesting

Manufacture

10


Final Product

• USP <61>• USP <62>• Bioburden

Testing

• USP <51>• Water activity• Container/Closure

Stability

11


Manufacturing Process Stream…

Think

…Think “holistically”

12


What is “Characterization”?

13


Characterization is represented by the establishment of microbial profiles, including the evaluation of sources, routes of ingress and susceptibility to elimination or reduction.

(Or… Which bugs are predominant, where are they and how can I control them?)

14


“Case Studies of Microbial Contamination in Biologic Product Manufacturing”Kalavati Suvarna, Ph.D, Anastasia Lolas, M.S., Patricia Hughes, Ph.D. and Richard L

Friedman, M.S.American Pharmaceutical Review, January/February 2011

15


Raw Materials/Component

s• Inherent bioburden• Test for objectionable organisms

Manufacture

• Environmental organisms (surface/air)

• Bioburden of utilities

• Exposure to human-borne organisms

• In-process bioburden

• Equipment cleaning• Open vs. closed

Final Product

• Bioburden• Objectionable organisms

EM ReportsClean Utility Reports

Annual Product Reviews



16


Some examples…


sNatural Mold, Spore-forming

bacteria


sAqueous Gram-negatives

Utilities Deionized Water Gram-negatives

Facility HVAC Airborne organisms

Manufacture Personnel Human-borne organisms

Manufacture In-Process Testing Predisposition to objectionables

17


Characterization…

Focus on the Types of Organisms

18


Why Characterize Your Microbial Isolates?

19


Risk• Assessment• Management• Mitigation

Implementation into Compendial and other Microbiological Tests

Two reasons…

20


TITLE 21--FOOD AND DRUGS CHAPTER I--FOOD AND DRUG ADMINISTRATIONDEPARTMENT OF HEALTH AND HUMAN SERVICES SUBCHAPTER C--DRUGS: GENERAL PART 211 -- CURRENT GOOD MANUFACTURING PRACTICE FOR FINISHED PHARMACEUTICALS Subpart F--Production and Process Controls Sec. 211.113 Control of microbiological contamination. (a) Appropriate written procedures, designed to prevent objectionable microorganisms in drug products not required to be sterile, shall be established and followed. (b) Appropriate written procedures, designed to prevent microbiological contamination of drug products purporting to be sterile, shall be established and followed. Such procedures shall include validation of all aseptic and sterilization processes.

21


“HACCP [Hazard Analysis Critical Control

Point] might be used to identify and manage

risks associated with physical, chemical, and

biological hazards (including microbiological

contamination).”ICH Q9: Quality Risk Management

U.S. Department of Health and Human ServicesFood and Drug Administration

Center for Drug Evaluation and Research (CDER)Center for Biologics Evaluation and Research

(CBER)June 2006

22


“Where microbiological specifications have been established for the intermediate or API, facilities should also be designed to limit exposure to objectionable microbiological contaminants, as appropriate.”

“If drinking (potable) water is insufficient to ensure API quality and tighter chemical and/or microbiological water quality specifications are called for, appropriate specifications for physical/chemical attributes, total microbial counts, objectionable organisms, and/or endotoxins should be established.”

“Appropriate specifications should be established for APIs in accordance with accepted standards and consistent with the manufacturing process. … If the API has a specification for microbiological purity, appropriate action limits for total microbial counts and objectionable organisms should be established and met.”

“Appropriate microbiological tests should be conducted on each batch of intermediate and API where microbial quality is specified.”ICH Q7A: Good Manufacturing Practice Guidance for Active Pharmaceutical

IngredientsU.S. Department of Health and Human Services

Food and Drug AdministrationCenter for Drug Evaluation and Research (CDER)

Center for Biologics Evaluation and Research (CBER)August 2001

23

“Characterizing microorganisms recovered from environmental and personnel monitoring is an important part of surveillances programs.”

“…routine characterization should continue to determine whether isolates are part of the normal microbial flora or represent something different.”

“A change in the microbial flora or the introduction of a previously undetected species might signify a change in a system that should be investigation.”

“Characterizations can be useful clues as to the possible source of isolates.”


Technical Report No. 13: Fundamentals of an Environmental Monitoring Program

Journal of Pharmaceutical Science and TechnologyParenteral Drug Association

September/October 2001, Supplement TR13, Vol. 55, No. 5

24


Regarding recovery of unspecified microorganisms…

“Where warranted, a risk-based assessment of the relevant factors is conducted by personnel with specialized training in microbiology and in the interpretation of microbiological data. For raw materials, the assessment takes account of the processing to which the product is subjected, the current technology of testing, and the availability of materials of the desired quality.”

USP 33 <1111> MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: ACCEPTANCE CRITERIA FOR PHARMACEUTICAL PREPARATIONS AND

SUBSTANCES FOR PHARMACEUTICAL USE

25


“A well-established microbial EM program must identify, at least at the genus level, microorganisms isolated from samples that reach the alert and action limits established. Evaluation of the isolates is necessary and would be evaluated for their ability to grow in the non-sterile product present at the time of sampling. It is necessary to make a list of microorganisms isolated and identified at least at the genus level during the first year of implementation. This list is crucial to identify the profile of microorganism species found in production areas, its possible origin, trends, and seasonal peaks.”

“Implementation of Microbial Environmental Monitoring Program for Non-Aseptic Pharmaceutical Processes”Angel L. Salamán-Byron, Ph.D.

Wyeth Pharmaceutical CompanyAmerican Pharmaceutical Review, July/August 2006

26


“Since the microbial quality of pharmaceutical products is a function of microorganisms introduced through excipients, the significance of microorganisms recovered from excipients should be evaluated in terms of the number and type of organisms, whether or not they will survive the manufacturing process, their ability to grow in the product formulation, the use of the product and the potential hazard to the user.”

“Microbiological Considerations When Selecting Excipients During Product Development”

Eda Ross Montgomery, Ph.D (Vertex Pharmaceuticals Company)and Wireko Manu-Tawiah, Ph.D (Wyeth Pharmaceuticals)

American Pharmaceutical Review, July/August 2004

27


“Which temperature do you think should be the minimum acceptable and for how long to assess that it has not had any impact on the viability of possible contaminants in the samples during shipments?”

LinkedIn Group: Pharmaceutical Microbiology ForumDolors G.

March 3, 2011

28


“It's depending on what your normal storage conditions are? Have you prove that your normal storage conditions are ok (microbiological wise). It's also depending on what your microbiological starting contamination is (before shipment), not only the amount but also the kind of micro-organism. Do you have any data about the temperature during transport.”

LinkedIn Group: Pharmaceutical Microbiology ForumEsther B.

March 4, 2011

“Ambient temperature 20-25ºC should be fine providing the material is packaged correctly to prevent moisture ingress. The preservation of solid dose forms [relies] on a low available water value (Aw), this analysis should be part of your risk assessment for storage and transporting a pharmaceutical preparation.”

LinkedIn Group: Pharmaceutical Microbiology ForumKarl B.

March 4, 2011

29


“For clarification, the question is … if any possible present microbe can die at low temperatures.”

LinkedIn Group: Pharmaceutical Microbiology ForumDolors G.

March 4, 2011

“The simple answer is yes. Organisms present in pharmaceutical preparations are usually in a stressed state and lower temperature can result in cell death. This does however depend on the organism and the matrix of the product.”

LinkedIn Group: Pharmaceutical Microbiology ForumKarl B.

March 4, 2011

“I'd not count on lowered temperature to address microbial contamination.”

LinkedIn Group: Pharmaceutical Microbiology ForumPhilip Geis

March 4, 2011

30


“Although drugmakers have begun to perform environmental monitoring in their nonsterile manufacturing facilities, many companies are uncertain about how to interpret the data and develop alert and action levels, says Leonard W. Mestrandrea, principal consultant at Mestrandrea Consulting and chair of the [USP Expert Committee on Microbiology]. The industry should record the types and numbers of microorganisms that it identifies in its environmental-monitoring programs and evaluate the information on a quarterly basis, he says. This strategy could provide guidance, based on scientific rationale, for assessing and controlling the risk of contamination.”



31


Interestingly, there is some debate. From the same article, Dr. James Akers is quoted…

“I don’t think a scientific reason for increased emphasis on nonsterile product monitoring has been established”

“The consensus among expert microbiologists is that microbiological environmental-control level recommendations would be of little benefit for the vast majority of nonsterile products”

“The focus should be on reducing risk in processing, not on bug hunting”



32


Implementation of In-House Isolates into Compendial Testing

THE DEBATE!

33

“I've previously posted feedback requests on the PMF regarding the use of in-house isolates [for compendial testing]. The responses were varied, but included an opinion that the use of these organisms is scientifically irrelevant. The reasoning included (a) EM isolates cease to become "environmental" once they've been cultured in the lab (the same could be said for ATCC organisms), and (b) it's too difficult to properly standardize and maintain them. As we all well know, "science" and "regulations" often clash… Should we try to argue the scientific irrelevancy of using in-house isolates, and risk receiving a 483, or should we just comply with regulatory expectations?”


Robert WestneyPMF List Posting. July 22, 2010

Why the debate?

34


1 mL

Microbial Suspension

9 mL saline

Plate 0.1 mL

TSA

Final Dilution Factor = 10-2

1 mL

9 mL saline

Plate 0.1 mL


1 mL

9 mL saline

Plate 0.1 mL


etc.

TSA TSA

Standardization of the microbial suspension (example)…

35


Implementation of In-House Isolates• Disinfectant Efficacy

• USP <51>, “Antimicrobial Effectiveness Testing”

• Media Growth Promotion

• USP <61>, “Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests” – Suitability Test

• USP <62>, “Microbiological Examination of Nonsterile Products: Tests for Specified Microorganisms” – Suitability Test

• Bioburden Method Qualification

• Container-Closure Integrity Testing (CCIT)

• Sterilizing filter studies

• Qualification of Rapid Microbiological Methods (RMM)

• Analyst Qualification

• Biological Indicators (USP <55>)

36


Disinfectant Efficacy

37


“… 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 33 <1072> DISINFECTANTS AND ANTISEPTICS

“To demonstrate the efficacy of a disinfectant within a pharmaceutical manufacturing environment, it may be deemed necessary to conduct the following tests: (1) use-dilution tests (screening disinfectants for their efficacy at various concentrations and contact times against a wide range of standard test organisms and environmental isolates); (2) surface challenge tests (using standard test microorganisms and microorganisms that are typical environmental isolates…”

38

“It is a sound practice to perform challenge testing of the selected sanitizers/disinfectants with isolates routinely recovered by the environmental monitoring program. This establishes the practical effectiveness of the disinfectants.”

Technical Report No. 13: Fundamentals of an Environmental Monitoring Program

Journal of Pharmaceutical Science and TechnologyParenteral Drug Association

September/October 2001, Supplement TR13, Vol. 55, No. 5

39


“If using an Association of Official Analytical Chemists (AOAC) method, microorganisms specified in the reference which are most likely to be found in the manufacturing environment should be used and tests performed on microbial isolates frequently found in the environment can provide additional information on the effectiveness of the disinfectants.” “Regulatory Aspects Concerning the Quality Controls of Microbiological

and Nonviable Particulate Contamination in Pharmaceutical Manufacturing”

Ray T. OjiFood and Drug Administration

American Pharmaceutical Review, January/February 2004

“Disinfectant validation involves the documented verification and implementation of procedures that have been shown to achieve adequate control over the range and levels of microorganisms that may be encountered in a facility.” “Disinfectant Validation”

Darren Wallis, GlaxoSmithKlineAmerican Pharmaceutical Review [unknown publish date]

40


FDA Warning Letter…

“Disinfectant effectiveness studies against representative microorganisms and/or specific in-house isolates were not conducted for cleaning agents used in your facility to disinfect production areas…” [March 29, 2008]

41


USP <51>, “Antimicrobial Effectiveness Testing”

42


USP <51> (Antimicrobial Effectiveness Testing) doesn’t mention the use of in-house isolates, but…

USP 33 <1227> VALIDATION OF MICROBIAL RECOVERY FROM PHARMACOPEIAL ARTICLES

“The use of membrane filtration to recover challenge microorganisms, or the use of environmental isolates as challenge microorganisms in antimicrobial effectiveness testing, requires validation of the countable range.”

43


“We suggest that the challenge microbes include the panel provided in USP<51>, as well as typical skin microflora and nosocomial agents to simulate the types of flora that may contaminate a drug product in a hospital pharmacy.”

“Microbiological Quality of Drug Products after Penetration of the Container System for Dose Preparation Prior to Patient Administration”

John W. Metcalfe, Ph.D. FDA/CDER/OPSAmerican Pharmaceutical Review, [unknown publish date]

44


“Use of organisms beyond that required by the compendia is also suggested…”

“Making Sense Out Of Microbiological Testing To Support Product Stability of Non-Sterile Pharmaceuticals”

Linda Skowronsky, GlaxoSmithKlineAmerican Pharmaceutical Review, [unknown publish date]

Regarding Antimicrobial Effectiveness Testing…

45


Media Growth Promotion

46


USP 33 <1117> MICROBIOLOGICAL BEST LABORATORY PRACTICES

“… microorganisms used in growth-promotion testing may be based on the manufacturer's recommendation for a particular medium, or may include representative environmental isolates (but these latter are not to be construed as compendial requirements).”

USP 33 <1116> MICROBIOLOGICAL EVALUATION OF CLEAN ROOMS AND OTHER CONTROLLED

ENVIRONMENTS

“… for the Growth Promotion test, representative microflora isolated from the controlled environment or ATCC strain preparations of these isolates may also be used to test media.”

47


Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing

Current Good Manufacturing PracticeU.S. Department of Health and Human Services

Food and Drug AdministrationCenter for Drug Evaluation and Research (CDER)

Center for Biologics Evaluation and Research (CBER)Office of Regulatory Affairs (ORA)

September 2004

“The QC laboratory should determine if USP indicator organisms sufficiently represent production-related isolates. Environmental monitoring and sterility test isolates can be substituted (as appropriate) or added to the growth promotion challenge.”

48


“...the firm does not test TSA...during growth promotion tests for microorganisms to include for example, molds, yeasts and other potential known environmental contaminants found in the manufacturing facility and/or raw materials.”

FDA 483 ObservationMcNeil Consumer Healthcare

Fort Washington, PAApril 30, 2010

49


Sofia G.PMFList posting

Apr 29, 2010

“Does anyone perform growth promotion with environmental isolates? If so, how do you manage such a program and are there any references for this requirement?”

50


“I would argue that as soon as an environmental isolate is put onto growth media, it is no longer an "environmental" isolate. Meaning that its growth characteristics have been altered by sub-culturing onto enriched media. I don't know what added value that brings to the table.”

Rick JakoberApril 29, 2010

“I tend to agree. However, the same can be said for type culture organisms. All things being equal, what's more relevant to a firm's media growth promotion regimen?... Demonstrating that a medium can recover (e.g.) an organism isolated from someone's outer ear infection 67 years ago (P. aeruginosa ATCC 9027), or an organism from the firm's own site? I would argue that the use of type culture organisms is completely irrelevant, compared to the use of in-house isolates.”

Robert WestneyApril 30, 2010

51


“While I fully agree with you that an environmental isolate, once culturedfor several passes in the lab is no longer the same bug, one of the veryfirst questions we are asked about our environmental program by almost every regulatory inspector is if we growth-promote with environmental isolates.

Another case of compliance vs science.”

Carole G.June 4, 2010

52


“I use recovered organisms, in addition to ATCC strains, regularly for growth promotion of media fills to ensure that contamination by those specific isolates during production would be detectable in the media.”

Gwendolyn H.April 30, 2010

“I agree with Mr. Jakober, the use [of an] environmental isolate does not add value to the growth promotion process. For growth promotion I am not aware of any requirement to utilize an environmental isolate.

I would also add the same argument could be said of the use of environmental isolates in disinfectant studies, however it seems to be the industry norm to use them in these studies.”

Jeff O.May 3, 2010

53


“The compendia recommends the use of specified strains for compendial tests as they need to be standardized.

Irrespective of the origin of these strain cultures, these QC organisms have a utility of a long history of use by both media manufacturers and lab users, the ability to source them from national culture collections or prepared commercial inocula derived from those cultures and are GPT requirements for media in general test methods.

Environmental isolates are often mismanaged by the microbiology lab.”

Tony CundellJune 4, 2010

54


“Given that most companies are using the Ph Eur requirement of 5 days at 30-35 degree C B.subtilis (spizizenii) ATCC 6633 and P.aeruginosa ATCC 9027 plus an environmental isolate like R. picketti may be a good choice.”

Tony CundellPMFList posting (regarding GP of media used for water testing)

February 2, 2010

“In general, I do not support the wholesale use of in-house cultures for growth-promotion testing.”

Tony CundellPMFList posting (regarding GP of media with EM isolates)

June 3, 2010

“Growth promotion should be demonstrated using organisms listed in USP General Chapter <71> as well as environmental, personnel, and sterility test failure isolates [at the] <100-cfu challenge.”

“Microbial Testing in Support of Aseptic Processing”Anthony M.Cundell

Pharmaceutical Technology, June 2004

55


“I've previously posted feedback requests on the PMF regarding the use of in-house isolates [for compendial testing]. The responses were varied, but included an opinion that the use of these organisms is scientifically irrelevant. The reasoning included (a) EM isolates cease to become "environmental" once they've been cultured in the lab (the same could be said for ATCC organisms), and (b) it's too difficult to properly standardize and maintain them. As we all well know, "science" and "regulations" often clash… Should we try to argue the scientific irrelevancy of using in-house isolates, and risk receiving a 483, or should we just comply with regulatory expectations?”

Robert WestneyPMF List Posting. July 22, 2010

56


“This is one of the problems with using 483 observations as "cGMP". They are not. They are the observations of one inspector who may or may not be knowledgeable in microbiology.

The company receiving the 483 observation, in turn, makes a business decision as to whether they will lose more money complying with a questionable request, or delaying approval by arguing. Usually the company goes along with the less onerous requests and a new ‘best practice’ is born.”

Scott SuttonPMF List Posting. July 22, 2010

57


“Each time I review our Growth Promotion SOP I enter into this same debate. Yes, we have set rules on enrolling isolates from the EM Program into our Growth Promotion panel, but I question the bacteriological merit of this practice.

Acceptable user compendial growth promotion tests are indicators that the culture media will recover a broad spectrum of microbial isolates. What then is the value add[ed] of the performance of a Staphylococcus or Bacillus species (for example) once recovered from our environment and propagated for release testing of culture media to our growth promotion programs?

We are responsible for managing myths and perceptions with sound science…”

Robert M.July 22, 2010

58


“As others have said, what you've got is an OBSERVATION- one inspector'sopinion. This does not make it a ‘Regulatory Expectation’… Personally, I don't think this point is a 'show-stopper' as a 483 observation- and many companies simply don't have the qualified equipment and personnel to preserve and maintain stock cultures of their environmental bugs.”

Michael C.July 23, 2010

“I would argue the science and the logic behind for not trying to recover EM organisms during Growth Promotion test. I totally agree that EM organism will stop to behave as such after a few transfers and will be nothing more then the purchased one.”

J.B.July 26, 2010

59


Microbiological Method Qualifications

60


• USP <61>, “Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests” – Suitability Test

• USP <62>, “Microbiological Examination of Nonsterile Products: Tests for Specified Microorganisms” – Suitability Test

• Bioburden Method Qualification

61


No mention in USP <61> or <62> of the use of in-house isolates for “Method Suitability” testing.

No mention in USP <1227> of the use of in-house isolates for “Method Suitability” testing (other than that previously cited for AET).

Is it still worth consideration?

62


“The author highly suggests performing test method suitability studies using wild-type isolates from production surfaces instead of laboratory-adapted strains of bacteria. Wild-type strains are a better representation of the organisms encountered on production areas than those strains that lack wild characteristics.”

“Bioburden Method Suitability for Cleaning and Sanitation Monitoring: How Far Do We Have to Go?”

Angel L. Salaman-ByronPharmaceutical Technology, August 2, 2010

63


• Qualification of Rapid Microbiological Methods (RMM)

• Sterilizing filter studies• Container-Closure Integrity Testing (CCIT)• Analyst Qualification• Biological Indicators (USP <55>)

64


“Validation studies evaluated sterility test samples from primary culture, expansion culture, and final product culture with a microbial challenge comprised of 10 microbial species. These species included a mix of commercial reference strains and stressed cells from frozen archives of environmental isolates and sterility test failures. During the validation process, FDA recommended challenge microorganisms that prompted additional method optimization resulting in improvements to incubation temperature and media formulation.”

“Automated Rapid Microbiological Methods for the Biopharmaceutical Industry: Selection, Validation, and Implementation for an Autologous Cell Therapy Product”

John Duguid & Gary C. du Moulin Ph.D. Quality Systems, GenzymeAmerican Pharmaceutical Review, [unknown publish date]

65


“No one organism type is likely to serve as a model for all in defining the “sterilizing filter.” B. diminuta filled its role rather well. Drug isolates would seem to be logical candidates as models in any particular case. For practical reasons, the universe of possible model organisms consists of those likely to be encountered in pharmaceutical settings, a group whose identity is not universally agreed upon.”

“The Sterilizing Filter and Its Pore Size Rating”

Theodore H. Meltzer, Ph.D. and Maik W. JornitzCapitola Consulting Co. and Sartorius North America

American Pharmaceutical Review, Fall 2003

66


CONCLUSION• Understand your “Manufacturing Process

Stream”

• Know how to “characterize” your in-house isolates

• Use the knowledge you gain from characterization of your in-house isolates to

o Assess, manage and mitigate risk

o Decide if it’s relevant to incorporate them into your microbiological testing

67


Use the knowledge you gained from characterization of your in-house microbial isolates…

68


Robert Westney, M.S., RAC, CMQ/OE

Principal Consultant

Westney & Associates Consulting, LLC

[email protected]

267-981-2419

Thank you!

69


Q & A

Documents

Characterization of Microbial Isolates from the Manufacturing Process Stream