Implementation of Good Laboratory Practice in a University Research Unit Xavier Abad1,*, Albert Bosch1 and Carme Navarro2 1
Enteric Virus Laboratory, Microbiology Department, School of Biology, University of Barcelona, Av. Diagonal, 645, 08028 Barcelona, Spain 2 Quality Assurance Unit, Scientific-Technical Services, University of Barcelona, C. Josep Samitier, 1-5, 08028 Barcelona, Spain
Summary This article describes our experience in the implementation of Good Laboratory Practice (GLP) in a research group of the Department of Microbiology in the University of Barcelona (UB). Some issues relating to quality assurance in a research laboratory setting are reviewed, and several comments and suggestions arise from our experience in setting up GLP. Copyright # 2005 John Wiley & Sons, Ltd. Key Words: GLP; Good Laboratory Practice; enteric viruses; virus safety; virus removal; inactivation
Introduction The Enteric Virus Laboratory (EVL) at the University of Barcelona has been conducting research on environmental virology since 1978, with particular attention to hepatitis A virus, rotaviruses and astroviruses, but also to other enteric viruses. One of our research areas is the study of viral inactivation and removal, and its mechanisms, and how to eliminate viruses from water, fomites, foods and other environments. We are also pursuing several other research areas, including molecular studies on hepatitis A virus, molecular epidemiology of rotaviruses and astroviruses, etc. All of these have been performed by two professors, some technical personnel and several PhD students; as a consequence more than one hundred international publications and hundreds of communications to international congresses and workshops have been produced. As our reputa*Correspondence to: F. X. Abad, Enteric Virus Laboratory, Microbiology Department, School of Biology, University of Barcelona, Av. Diagonal, 645, 08028 Barcelona, Spain. E-mail:
[email protected] Copyright r 2005 John Wiley & Sons, Ltd.
tion in the field of viral inactivation grew, several private companies, forced by regulations to certify the capability of their processes for the inactivation or removal of viruses, contacted us with requests to validate their methods for viral elimination. The evaluations needed to be conducted in compliance with Good Laboratory Practice (GLP) principles. This article describes our experience in implementing GLP in our own Viral Validation Unit (VVU) within the EVL. This implementation achieved GLP compliance certification in 2001. An historical outline of this implementation is shown in Table 1.
Objectives Our goals were to provide reliable, high quality analytical data to support the research and development efforts of our industrial sponsors. GLP are the recognized rules governing the conduct of non-clinical safety studies. They ensure the quality, integrity and reliability of the study data; they reduce the likelihood that Qual Assur J 2005; 9, 304–311. DOI: 10.1002/qaj.352
Implementation of GLP in a University Research Unit
Table 1. History of GLP Implementation in the EVL Year
Activity
1996
Writing SOPs for the most frequent experimental procedures and apparatus Implementation of study inspections from the QAU First Facilities Inspection from the QAU Move to facilities in a new research building Settlement of final EVL personnel organizational chart and decision flow diagram Achievement of GLP certification Most recent renewal of GLP compliance
1998 1998 2000 2000
2001 2003
our industrial sponsors will need to spend money on repeating any stage of these experiments and they provide the basis for a good working relationship with our sponsors. We also recognized that achieving GLP status, regardless of its workload, would be of benefit to our own research, for example developing standard operating procedures (SOPs), development of standard controls, and instrument calibration.
Organization All organizations have their own ‘natural’ quality systems, though often they are informal systems which do not follow a specific standard (e.g. GLP). As such, some of the road has already been traveled in any organization prior to any decision to implement GLP. To benefit from the collective experience in the EVL, our first activity was to identify and formalize the elements of our expertise and then to focus on how best they could be used to meet the goals of our industrial sponsors. Research is a continuously evolving process aimed at discovering new facts. By its own nature, the research work of PhD students is subjected to changes in direction in response to new and often unexpected results. In fact, the end-result of research may be unrelated to the initial aims of a research project. Moreover, in our field, short-term contracts are common practice leading to high staff turnover. Whilst this can bring vigor to a research group, it can Copyright r 2005 John Wiley & Sons, Ltd.
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also detract from establishing the mechanisms that ensure consistency and reliability of data over the long term. For these reasons, and because quality assurance (QA) can be more readily implemented in research and development than in basic research [1], we decided to create a VVU, within the EVL, specialized in methods and techniques for external sponsors, eventually leading to a ‘routine research’ activity, in our case virus validation studies with its own regulations to become GLP compliant [2–6]. The advantage of designating a separate study unit is that it could respond to the demands of our industrial sponsors, and at the same time not detract from the atmosphere of flexibility and freedom in other areas of basic research in our laboratory. The organization of the VVU within the EVL is described in Figure 1. Firstly, a Head of Laboratory (HL) (the vertex of the pyramid) was appointed, who would be ultimately responsible for implementation of GLP. Secondly, two Study Directors (SD) were appointed, contracted as full-time university staff, and highly skilled in virology. At the initiation of each study, a SD is assigned and this person takes responsibility for GLP compliance, being the point of contact for VVU staff on that study and for the external collaborator. The usual tasks of a SD are: *
*
*
Generate a study protocol in response to a request from an outside sponsor. Assume responsibility for the overall conduct of the study, particularly all decisions that may affect the integrity of the study. Generate the final report that is sent to the study sponsor.
In addition to these normal duties, one SD was also designated by the HL as the GLP coordinator, to be responsible for implementing the principles of GLP and for compliance within VVU. The tasks of this GLP coordinator are: *
*
Generate accurate job descriptions, and a clear delineation of the decision flow path. Write SOPs concerning the use of apparatus, sub culturing of cell lines, production of viral stocks, etc. Qual Assur J 2005; 9, 304–311.
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QUALITY ASSURANCE UNIT (QAU)
HEAD OF LABORATORY
Head of QAU and other personnel Function: to audit viral validation studies and control compliance of GLP principles
VIRAL VALIDATION UNIT
RESEARCHS UNITS Research staff, Pre- and Post-doctoral students Function: Basic research and development of techniques
Study directors Other personnel Function: to develop and conduct GLP-compliant viral validation studies
TECHNICAL STAFF Function: supply media and reagents for research units and for viral validation unit.
Figure 1. Organizational chart of the EVL
*
Create data systems to record the arrival, maintenance, use and destruction of cell lines and viral strains.
We also needed a system for independent evaluation of all activities within the VVU. This was achieved by setting up a QAU that serves the entire University, and is located within the Scientific Technical Services. The QAU personnel are highly experienced in implementing GLP and ISO standards, are not funded by our group, and operate under a different hierarchical chain. The QAU personnel carry out the following activities: *
*
*
Verify protocols in order to assure their compliance with GLP principles. Conduct inspections of facilities and studies, and report their results to the SD and HL. Audit final reports, prepare and sign a quality assurance statement.
In addition to day-to-day training given by SDs, all staff within the EVL (whether associated with the VVU or not) receive at least one day a year of GLP training given by the QAU. Copyright r 2005 John Wiley & Sons, Ltd.
Training records for all personnel are maintained by QAU.
Facilities The test facilities occupy nearly 40 square meters. Additionally, an independent room for performing validation experiments is available when complex procedures or special environmental conditions (temperature) require a second area. Samples are received and processed in a clean area well-segregated from the validation room. This area is provided with several airflow cabinets and a biohazard cabinet. Separate areas have been designated for the maintenance of uninfected cell lines used to test and grow the viruses, and for the viruses themselves with infected cell lines (virus/cells couple, also called experimental systems). All of these precautions ensure a proper degree of separation of the different activities in each study and help prevent contamination and mix-ups upon receipt, testing and storage of materials. A ‘transient’ archive facility has been set up within Qual Assur J 2005; 9, 304–311.
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the EVL for the safe storage and retrieval of study protocols, raw data notebooks, sponsor and SD communications, and final reports. The ultimate destination of these records is the storage facilities in the QAU. Waste collection, storage and disposal have been designed in such a way as not to jeopardize the integrity of the studies nor the health and safety of other staff in the Department of Microbiology. All biological systems, infected or not, are autoclaved at the appropriate temperature and time period.
Apparatus, Material, and Reagents The apparatus used in the studies are periodically inspected, cleaned, maintained, and calibrated according to internal SOPs. Records of all these activities are maintained and transferred after each test facility inspection to QAU. All chemicals, reagents and solutions (PBS, trypsin solutions, etc.) are labeled to indicate identity (with concentration if appropriate), expiration date and specific storage instructions. Additional information is available for each reagent and solution as a separate written record describing the source (suppliers, batch identification), preparation date, person in charge and stability.
Biological Experimental Systems (also called ‘test systems’ in the OECD GLP Principles [7]) For some authors [8] GLP standards define the test system as ‘any animal, plant, microorganism, or subparts thereof to which the test or control item is administered or added for study’. By this definition, some authors [9] consider the best technical analogy to label as test system the scaled-down manufacturing process and the product intermediate as the test article. Our aim is to measure the efficacy of viral elimination or decay in a manufacturing process. We have defined the test item as both the intermediate from a manufacturing process, and/or the actual step, scaled down from the manufacturing process, that is intended to remove Copyright r 2005 John Wiley & Sons, Ltd.
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viruses. The test system is the couple virus plus cell line that responds to the test item with a change in viral titer (Figures 2(a) and 2(b)). By its very nature, the use of a binomial test system with two biological components is potentially subject to high variability from study to study. Therefore, we perform robustness assays for each experimental (test) system to ensure that changes in post-infection media composition, adsorption time, age of host cells, or differences in cells passage number have not affected final viral titer. Records of source, date and condition of arrival of experimental test systems (viral strains and cell lines) have been maintained since 1994. All information needed to properly identify the test systems appears on their container (usually plastic micro plate or flasks): cell line name, growth medium, date of subculture, and operator are noted for non-infected cell lines. Infected cell lines are additionally noted with virus strain, sample code, assayed dilutions, protocol code, and operator. Containers are of single-use, consequently when data (virus titers) are recorded all these materials (both infected and non-infected) are destroyed by autoclaving at appropriate temperature and time period.
Test and Reference Items Upon receipt of the test item, records are started which note the sample characteristics, date of receipt, expiration date, recipient, and quantities. A sample is taken to verify that sample characteristics match those reported to us; and appropriate records are kept (date of extraction, quantity withdrawn, remaining weight, study code, and responsible person) each time a sample is withdrawn. Storage containers carry identification information, expiration date, and specific storage instructions. According to GLP regulations (No. 1 Series OECD [7]), each test item has to be properly identified with code number, chemical abstracts service registry number, name, and biological parameters. In many viral validation studies, however, this Qual Assur J 2005; 9, 304–311.
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Figure 2. (a) Typical relationship between test system and test item; and (b) relationship between test system and test item in viral validation studies. If characteristics of product intermediate or critical parameters of manufacturing steps are changed, the viral persistence could change, too. So, new validation studies should be performed. *The viral suspension alone cannot be the test system since we need a cell line for replication of the virus in order to evaluate its infectious titer. It is the combination of virus strain and susceptible cell line that form the test system (or experimental system). Moreover, results differ using same virus strain and several susceptible cell lines, or using quite similar viral strain and the same susceptible cell line.
detailed characterization is impossible, as we work with intermediates produced from industrial processes with only a few biologically or chemically defined parameters. Moreover, not only the intermediate, but also the inactivation step itself can be considered as test items, if we take into account than we are evaluating the Copyright r 2005 John Wiley & Sons, Ltd.
response of an experimental system to a defined inactivation treatment, when diluted in a matrix (intermediate) (Figures 2(a) and 2(b)). We require sponsors to provide the identity, batch number, composition data, concentrations, and other defined characteristics appropriate to define each batch of test item. Qual Assur J 2005; 9, 304–311.
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Standard Operating Procedures SOPs provide documentation of all routine experiments. Strict adherence to the SOPs ensures the quality and integrity of data generated, and allows comparison of results from different experiments. In our test facilities more than one hundred SOPs (from use of apparatus to experimental processes) have been written and subsequently approved by the Head of Laboratory. SOPs are reviewed once every two years, with up to two revisions, or a period no longer than six years before a new SOP must be generated and approved. QAU personnel coordinate all revisions, distribute copies, and retain originals. All original operation manuals that may be used as supplements to SOPs are filed in a single place inside the laboratory to make consultation easy. Any deviation from an SOP is documented in each study notebook and acknowledged by the SD, who evaluates the necessity and impact of these deviations in relation to the whole study. Our SOPs cover the following main areas: 1.
2.
3.
4.
5.
Test items, reference items and starting materials: Receipt, identification, labeling, handling, and storage. Apparatus, materials and reagents: Use, maintenance, cleaning and calibration of apparatus; and preparation and labeling of reagents and solutions. Record keeping: Reports, storage and retrieval, coding of studies, data collection, indexing systems, and handling of data. Experimental test system (cell lines and viral strains): Procedures for receipt, proper placement, characterization, identification, care, preparation for a study, and handling of infected systems during the study. SOPs in groups 1–4 were written and reviewed by EVL personnel and approved by HL. In addition we comply with SOPs written and approved by QAA personnel detailing. Quality assurance procedures: Planning, scheduling, performing, documenting and reporting inspections by QAU personnel.
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All SOPs are distributed by the head of the QAU.
Performance of the Study The study protocol is the key document of a GLP study [10] since it describes the study that is to take place. For each study, the SD prepares a draft protocol and sends it to the study sponsor for comments. On return, the protocol is revised and sent to QAU for verification of GLP compliance. The final version of the protocol is approved, by dated signature by the SD and HL, verified for GLP compliance and signed by QAU personnel, and accepted by sponsor. Standard study protocol format is followed, according to GLP regulations [7] and the EURACHEM CITAC guide [11], and a final addendum of scientific and technical references and SOPs is included. After signing the protocol, any amendments are justified and approved by the SD, HL and study sponsors, and signed and dated by the SD. Deviations from the protocol during a study are described, explained, acknowledged and dated by the SD whose responsibility it is to evaluate the impact of the change on the integrity of the study. All deviations are also noted in the study raw data notebook and included in the final report. Besides the protocol, a strictly defined plan is distributed to QAU, HL and sponsor, indicating dates and hours of each experiment, with special attention to the critical steps of the process. We tend to receive two types of study proposals relating the evaluation of efficacy of particular steps in removal or inactivation of viruses from an industrial manufacturing process, or to ascertain the antiviral capacity of a defined formulation: 1.
The sponsor who is interested in evaluation of virucidal capability of a step in the production process, performs test runs of their manufacturing process using their own personnel, materials and apparatus. In this case, since the sponsor personnel performs the whole scale down industrial process, only sample analysis is within the Qual Assur J 2005; 9, 304–311.
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2.
framework of the GLP compliant facilities at EVL. The sponsor requests a study of the antiviral capacity of a defined formulation, and the inactivation assays as well as the analyses are performed by EVL personnel. In which case, all processes are under GLP compliance at the EVL.
Reporting of study results The SD prepares a final report containing the results and interpretation of raw data from the study notebooks. A draft report is sent to the sponsor for comments. After the addition of any relevant comments, a final draft is sent to QAU for auditing along with all other records and study notebooks. The final version of the report includes the statement of GLP compliance from the SD and the QAU statement, which list the inspections performed and their dates.
Archive of documentation The archive is located in the QAU facilities; it contains all the original documents relating to the studies performed by EVL under GLP principles, including study notebooks and other raw data. Access to the archive is restricted to personnel authorized by the HL, and documents cannot be taken out the archive without the permission of the sponsor.
Discussion and Conclusions Setting up a GLP system implies long-term commitment, not only by management, but also (and more importantly) by every staff member in the laboratory. A process that starts with the main goal of obtaining a compliance declaration or certification, rather than achieving real quality improvement, will be poorly accepted and certainly misunderstood by technical and scientific staff [10]. In our case, we feel that our GLP implementation has been successful for several reasons: probably the most important was that the system was developed ‘bottom–up’ Copyright r 2005 John Wiley & Sons, Ltd.
involving staff at all levels, rather than the ‘top– down’ approach of managerial imposition [12]. Secondly, it is the good science and technical competence of staff within the EVL and QAU, hand-in-hand with GLP principles, which brought us to achieve compliance [10,13]. GLP principles mainly apply to the formal aspects of a study (planning, performance and record keeping) and do not evaluate the technical competence of the study staff [10]. In fact, it was this technical competence that led our study sponsors to contact the EVL in the first place, and this remained the defining quality as we sought to apply GLP in the laboratory. This same principle was elegantly stated by Dent [13]; application of basic common sense and good science can bring us to find that we achieved compliance. A third factor in our favor was the small size of our group and well-defined management structure that allowed us to overcome some of the GLP implementation challenges that can prevail in universities [1,14]. Some challenges remain. Now that we have established GLP to comply with the requirements of our sponsors, we find that the principles of GLP are bringing benefit to other areas of research in the laboratory. Our most important challenge now is how to enhance our non-GLP activities with the best of the GLP principles. An obvious benefit that we have seen is that common experimental controls have been established, and laboratory instruments are maintained to a high working standard. A second benefit is that the GLP-compliant SOPs allow us to perform all research activities (GLP and non-GLP) under their scope in the same way. This brings us closer to ensuring reproducibility within our laboratory, and they have become an excellent training tool for laboratory workers. A third benefit that we have begun to see is that the principle of rigorous documentation in a more formalized manner results in good record keeping amongst all the students in the laboratory. Notebook keeping is so often at the whim of the individual and lacks uniformity, but this benefit of the GLP principles is seen by everybody. Qual Assur J 2005; 9, 304–311.
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Our next challenge is to develop a wider and deeper quality assurance system that will take into account good scientific and technical performance. This is not an unrealistic desire, but almost an obligation. In the same way as productive and service industries and public authorities take the principle of total quality management seriously, subcontractors such as university laboratories should also follow these principles. Although it is widely accepted that excellence in research is mostly located in universities and research institutes, in our opinion ‘excellence’ can no longer be simply based on reputation and promises, but on a well defined quality system, preferably certified or accredited. This system needs to be simple and flexible, to provide added value to the organization, and its implementation should not be considered as an end, but as a means to achieve higher technical standards within the setting of the laboratory research. In the near future, there may be no other alternative than to adopt some kind of quality assessment system, mainly due to external pressures from industrial contractors or public authorities. GLP principles are probably the most useful way to begin to fulfill this requirement.
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