establishing a glp compliance program for non-toxicology safety studies

Establishing a GLP Compliance Program For Non-Toxicology Safety Studies

Henry Li1,*, Susan Hawlk2, Hilton Renfrow1, Randy Hartwell1, Shih-Fong Chao1, GarrethSharp1, Connie Pilkington1, Steve Petteway, Jr.1, Kathryn Remington1 and Dominique Pifat1

1Bayer HealthCare LLC, Biological Products Division, 85 TW Alexander Drive, Research Triangle Park,NC 27709, USA2QA Consultant, 4969 Thornwood Trace, Acworth, GA 30102, USA

Copyright © 2004 John Wiley & Sons, Ltd. Qual Assur J 2004; 88, 94–101. DOI: 10.1002/qaj.270

Introduction

Many human plasma-derived and biotechnol-ogy products are life-saving medicines.

They, however, present the unique safety issue ofpotential viral contamination. This contamina-tion could occur through source plasma that con-tains pathogenic viruses or the introduction ofadventitious viruses during manufacture. To pre-vent contamination of biological products, manu-facturers have integrated a number of measuresinto the production processes. These includecareful selection and screening of source mate-rials and demonstration that manufacturingprocesses have sufficient capacity to remove orinactivate relevant and model viruses [1–4].

*Correspondence to: Henry Li, Bayer HealthCare LLC, BiologicalProducts Division, 85 TW Alexander Drive, Research TrianglePark, NC 27709, USA. E-mail: [email protected]

Summary

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Viral validation studies are performed todemonstrate the potential of manufacturingprocesses to clear adventitious viral contami-nants. To ensure the integrity of viral validationstudies, and thus ensure the viral safety of bio-logical products, regulatory agencies worldwidehave required these studies to be performed inaccordance with the principles of the GoodLaboratory Practices (GLP) [1]. BayerHealthCare manufactures biological products ofhuman plasma origin and a biotechnology prod-uct derived from cell culture. Recently, a compre-hensive GLP compliance program wasestablished for viral validation studies relating tothese products. Viral validation studies are dif-ferent from the traditional toxicology safety stud-ies for which GLPs were originally promulgated,and they presented some unique challenges inapplying GLP regulations.1 While the impetus forestablishment of our GLP program was primarilyviral validations, it was also important to makethe compliance program comprehensive enoughto meet the needs of other groups within Researchand Development, such as BioAnalytics andToxicology. Described here is our approach tomeet the challenges of implementing GLP compli-ance into non-toxicology safety studies for biolog-ical products.

Non-Toxicology SafetyStudies for Plasma-Derivedand Recombinant BiologicalProducts

A. Pathogen safety requirementsfor biological products

To ensure the quality of the source plasma, rigor-ous screening is done before its use in production.Plasma donors are screened to ensure goodhealth and the donated plasma is tested forknown clinically significant viruses, such ashuman immunodeficiency virus, hepatitis B virusand hepatitis C virus. Plasma testing occurs at

two levels; individual plasma units, in mini-pools,are tested and just prior to manufacture, produc-tion pools are tested. For the recombinant prod-uct, the source material is cell culturesupernatant fluid. Since cell lines may carryviruses, they are well characterized before use toidentify possible viral contaminants. In addition,raw materials, process intermediates and finalbulks are tested for adventitious viruses. Testingof source materials for plasma-derived andrecombinant products is inherent to the manu-facturing process, and as such, is governed byCurrent Good Manufacturing Practicies(cGMPs).

As a complement to screening source materials,the manufacturing processes for plasma-derivedand recombinant biological products must havethe capacity to remove or inactivate known oremergent viral pathogens [1–4]. The purificationprocesses for these proteins may have dual func-tions: purifying active protein components fromplasma or cell culture supernatant and removingor inactivating viruses. Often, dedicated viralremoval or inactivation steps are incorporatedinto the production streams to provide additionalviral clearance capacity [5,6]. The overall viralclearance capacity of the manufacturing processfor a biological product must be sufficient androbust, which is particularly important in light ofthe recent resurgence of infectious agents such asWest Nile virus, monkeypox virus, and the emer-gence of new viruses such as the severe acute res-piratory syndrome (SARS) virus [7]. Studies tovalidate virus clearance are in support of thesafety of the product and must be performedusing a small-scale model of the manufacturingprocess and in compliance with GLPs.

B. Viral validation studies support regulatory submissions

Data from viral validation studies are used to sup-port the viral safety of products at all stages ofdevelopment (Figure 1). When a biological thera-peutic is developed, the manufacturing processmust have a sufficient viral clearance profile,demonstrated by performing viral validationstudies. These viral safety results are included in

Copyright © 2004 John Wiley & Sons, Ltd. Qual Assur J 2004; 88, 94–101.

Establishing a GLP Compliance Program For Non-Toxicology Safety Studies 95

1Food and Drug Administration Good Laboratory Practice forNonclinical Laboratory Studies (GLPs).

96 Henry Li et al.


the Investigational New Drug (IND) application.Furthermore, viral validation studies are con-ducted to demonstrate the virus clearance poten-tial of the finalized manufacturing process. Theresults are included in the biological license appli-cation (BLA) to gain regulatory approval of theproduct. When a change is proposed in an existingmanufacturing process, the impact of the changeon the virus clearance potential of the process willbe determined by performing additional viral val-idation studies. The data obtained from thesestudies are submitted in Supplemental Changes tothe regulatory agencies. The application of GLPsto these studies ensures a standard of quality.

C. Validation requirements forconducting viral validation studies

Due to cGMP constraints as well as safety andpractical limitations in producing large quantitiesof infectious virus, validation of the virus clear-ance potential of a manufacturing process cannotbe conducted in the manufacturing facility. Inpractice, a bench-scale model of the manufactur-ing process is utilized in a laboratory suited forvirology studies.

An example of scaling down an addition andmixing step is shown in Figure 2. A 2500-l bulk

tank in the production process is scaled down5000-fold to a 0.5-l kettle at the bench level(Figure 2). Product intermediate is processedthrough the scaled-down step, matching all oper-ational parameters to production scale. Thevalidity of the scaled-down process is verified inthe laboratory by ensuring that biochemically,the product intermediates generated at small-scale meet pre-determined acceptance criteriathat are based on biochemical data from corre-sponding intermediates generated with the pro-duction-scale process.

To determine the virus clearance potential of agiven production step, infectious virus is spikedinto process intermediate and the processing stepis carried out using the validated small-scalemodel. Infectious virus is quantitated in thespiked intermediate prior to processing, as well asin the resulting fractions after processing. Thesemeasurements are used to calculate the virusreduction factor across the processing step. Thereduction in total viral load indicates the capacityof the step to remove or inactivate a given virus orfamily of viruses. The virus clearance factors forrelevant individual steps of a process are addedtogether to determine the overall virus clearancepotential for the entire manufacturing process.

An example of a scaled down nanofiltrationstep is shown in Figure 3. Nanofiltration can be

Figure 1. Viral validation studies in supporting regulatory submissions

an effective and robust viral removal step and isincreasingly incorporated in manufacturingprocesses to remove viruses. Nanofilters areavailable with different pore sizes and aredesigned to allow the passage of products, but notthe virus particles. In this example, the productintermediate, eluate from a chromatography col-umn, is spiked with an aliquot of a virus stock.Mimicking the flux used at the large scale, thespiked eluate is filtered through the nanofilterand the filtrate is collected. The differencebetween the virus load in the feed and in the fil-trate is used to calculate the viral reduction fac-tor for the nanofiltration step.

D. GLP compliance requirementsfor conducting viral validationstudies

Viral validation studies ensure viral safety forplasma-derived and recombinant products.Although it is a requirement that these studies beperformed in compliance with GLP regulations[1], it is also a good business practice to follow theGLP regulations when performing these expen-sive and complex non-toxicology studies. Datafrom these studies are typically used to supportproduct registrations and other regulatory sub-missions, sometimes several years after a study isconducted, and GLPs provide an added assur-ance of the integrity of the data.

Implementation of a GLPCompliance Program

A. Organization of the GLPprogram

Scientists in Bayer’s R&D group conduct a vari-ety of research and other scientific studies, manyof which are used to support regulatory submis-sions for new or existing products. For example,the Pathogen Safety group performs virus andprion clearance validation studies (non-toxicol-ogy safety studies), while the BioAnalytics groupdevelops and validates analytical assays used tocharacterize new and current products and inter-mediates. The Pharmacology group is responsiblefor performing pharmacology studies and coordi-nating toxicology studies. The goal in the estab-lishment of a GLP compliance program was toprovide uniformity, yet maintain the potential toaccommodate the different types of regulatednon-clinical studies that would be conducted bydifferent departments.

B. GLP training

One of the first steps in development of a GLPcompliance program was to achieve broad aware-ness of GLP regulations by implementing annual,site-wide intensive GLP training for employees inR&D, Quality Assurance (QA), Quality Control



Figure 2. Schematic diagram of a scaled-down production process

(QC) and Regulatory Affairs. During this one-dayintensive training, GLP history is introduced andthe GLPs, including 21 Code of FederalRegulations Part 58 and its preamble as well asthe Organization for Economic Co-operation andDevelopment (OECD) GLPs are covered. Sometopics are tailored to the group. For example, foremployees who have been primarily involved inmanufacturing, the differences between GLPsand cGMPs are discussed. Examples of FDAactions such as 483s, establishment inspectionreports (EIR) and warning letters are alsoincluded in the training. Annual GLP ‘refresher’sessions are conducted for personnel who havealready received the general GLP training.Besides general GLP training, specialized train-ing sessions are also provided to address specificneeds in GLP compliance, such as computer vali-dation and 21 Code of Federal Regulations (CFR)Part 11 compliance.

C. The good laboratory practicecompliance committee

One important aspect of the uniform GLP com-pliance program is the preparation of commonstandard operating procedures (SOPs). Thispractice ensures the uniform application of GLPstandards. To fulfill the requirement for a GLP-compliant study protocol, for example, an SOPwas written that standardizes the format of pro-tocols and amendments for all GLP-compliantstudies. To facilitate the harmonization of SOPsand GLP implementation across different groups,the GLP Compliance Committee (GLPCC), across-functional team, was formed to review allSOPs and to coordinate GLP activities. TheGLPCC consists of members from R&D and theQA Unit (QAU); Pathogen Safety, BioAnalytics,Pharmacology, and Preclinical Research repre-sentatives are part of the Bayer GLPCC, alongwith a representative from the QAU.

Every effort is made to keep abreast of newdevelopments in regulations. The GLPCC sub-scribes to compliance newsletters and journals,and holds regular meetings to discuss the changesin regulations and compliance audit trends. Also,the GLPCC members attend local and national

conferences. Computer validation and 21 CFRPart 11 compliance are areas in which regulatoryinterpretations of regulations have undergonerecent changes. To fully understand the currentregulatory thinking and the potential impact onGLP compliance, the GLPCC organized a one-day symposium. At this symposium, experts fromdifferent departments, such as RegulatoryAffairs, QA, Information Services, and R&D dis-cussed the FDA 21 CFR Part 11 regulation,‘Guidance for Industry, Part 11, ElectronicRecords; Electronic Signatures – Scope andApplication (FDA)’ that was issued in August2003, as well as other guidance documents oncomputer validation.

D. Application of GLP regulations to non-toxicologysafety studies

One of the challenges in applying GLP regula-tions to viral validation studies is that many GLPelements, such as the definitions for test articleand test system, do not directly apply to a scaled-down production process and virus clearanceexperiments. However, GLP elements are imple-mented whenever possible to ensure the integrityof the viral validation studies. The collaborativeefforts between QAU and R&D play an importantrole in interpreting GLP regulations and applyingGLP principles to these non-toxicology safetystudies. Both QAU and scientists from R&D care-fully evaluate viral validation studies for GLPcompliance.

In the GLP regulations, the term ‘test system’refers to ‘any animal, plant, microorganism, orsubparts thereof to which the test or control articleis administered or added for study’ [5]. Based onthe best technical analogy to the definition of thetest system in the GLPs, a scaled-down manufac-turing process in a viral validation study is definedas a test system (Figure 3). The product intermedi-ate to be processed by this manufacturing step isthen defined as a test article. Using the scaled-down nanofiltration step, shown in Figure 3, thetest article, the chromatography column eluate, isfiltered (administered to) through the nanofilter(test system) to generate filtrates (samples).

98 Henry Li et al.


Applying GLP definitions and principles toviral validation studies ensures the integrity ofthe study. Procedures for the proper handlingand storage of product intermediates (test arti-cles) according to GLP requirements have beenestablished. In addition, the receipt and use oftest articles are documented, according to GLPs.The implementation of GLP definitions and ele-ments for the study protocol, final report, devia-tion reporting and evaluation ensures that thestudy is properly conducted, documented, andaccurately reported. Management assigns a studydirector to each GLP-compliant study. A studyprotocol is prepared for each study, and isapproved by the study director and management.Trained study personnel then execute the studyprotocol. Any protocol deviations are docu-mented and reported to the study director in atimely manner. A final report is written andapproved. Procedures for uniform implementa-tion of these GLP elements have been delineatedin appropriate SOPs.

E. SOP Preparation, harmoniza-tion, distribution, and review

SOPs are an essential component of implementa-tion of GLPs [8]. The GLPCC is responsible forpreparing, reviewing, and harmonizing SOPs

used in regulated non-clinical laboratory studiesand sets a SOP preparation and review goal eachyear. When the GLP compliance program wasestablished, the GLPCC team, in conjunctionwith management, determined the critical SOPsrequired for the site GLP compliance program.SOP preparation was divided into phases. Thecritical, administrative SOPs, designed to estab-lish a framework for the GLP compliance pro-gram, were written as part of the first phase.Subsequent phases saw the generation of equip-ment and assay SOPs.

During SOP generation, drafts are reviewed bypersonnel from the various departments, fol-lowed by a review by GLPCC members as a team.Once an SOP is found to be acceptable, the final-ized version is submitted for QAU and manage-ment approval. The SOP preparation process,although time consuming, has become more effi-cient as the SOP authors become familiar withGLP regulations and the QAU has developed abetter understanding of the nature of these non-toxicology safety studies.

The GLPCC also coordinates GLP SOP train-ing to ensure all personnel are trained withrespect to relevant SOPs and that the execution ofSOPs is consistent. The GLPCC is also responsi-ble for coordinating an annual review of GLPSOPs to ensure that the SOPs are technically



Figure 3. Implementing GLP concepts to a scaled-down nanofiltration step

sound and are in compliance with the GLPregulations.

F. QAU

The QAU is independent of R&D and performsinternal and external facility audits for GLP com-pliance. In addition, QAU audits study protocols,final reports, and in-process phases of studies.The audit findings and observations are submit-ted to the study director and to management forreview and appropriate corrective actions. AQAU statement is issued for inclusion in eachfinal report.

QAU is represented on the GLPCC team,which allows for effective interactions betweenR&D and the QAU. This ensures that the QAU isaware of upcoming R&D activities. In addition,the QAU plays an important role in assisting inthe implementation of strategies for GLP compli-ance. A good working relationship has been estab-lished between the groups, such that input fromthe QAU on GLP compliance issues is activelysolicited by R&D.

When a study is contracted to a contractresearch organization (CRO) or contract labora-tory, GLP-QAU performs an assessment of theoverall regulatory compliance status of the CROor lab. Sometimes, scientists from R&D willaccompany the QAU auditor(s) on the audit inorder to evaluate the scientific capacity of thecontractor. An audit report will be submitted tothe contractor management and the appropriateBayer scientific/research area contracting thework. R&D will be responsible for the decision touse the contractor. A study monitor will be desig-nated to oversee the contracted study.

G. Archiving and disaster recovery

Proper management and archiving of raw dataand study-related materials are also very impor-tant to maintenance of GLP compliance. A desig-nated archivist is responsible for the storage ofcompleted study files. All study files are perma-nently archived at a contract archival facility thatmaintains current standards of GLP compliance.

To further safeguard raw data, once a study iscompleted, and before placing in the long-termarchive, study files are scanned into PDF formatand placed on electronic storage media, such asCDs. Scanned documents are visually verifiedand indexed for easy retrieval. After scanning,the study files are transferred to the approvedarchival facility for long-term storage. One set ofCDs containing scanned files is stored at a con-tract electronic media archival facility for disas-ter recovery.

Summary and Conclusion

Regulatory agencies worldwide require non-toxi-cology safety studies, such as viral validationstudies, to be conducted in compliance with theprinciples of GLPs. Establishment of a GLP com-pliance program requires collaboration amongmany groups. Furthermore, for non-toxicologysafety studies, there are additional challenges,such as applying elements meant for toxicologystudies to a non-toxicology system. A teamapproach has been used to adapt GLP require-ments, which were originally set forth for toxicol-ogy studies, to less conventional applications.

A key to the successful implementation of sucha GLP program is education regarding GLPs andto this end, general GLP training is conductedannually and SOP training is performed on a reg-ular basis. A cross-functional team plays animportant role in facilitating GLP complianceand in leading the efforts in preparation and har-monizing SOPs and other procedures to be usedby all groups within R&D when conducting orsupporting GLP-compliant studies. R&D scien-tists and the GLP-QAU continually work toimprove and expand the GLP compliance pro-gram for non-toxicology safety studies as well astoxicology studies at Bayer Biological Products.

Acknowledgements

Authors would like to thank GLPCC members,Stefan Burde, Proveen Dass, Mark Endsley, NehaFrantz, Amy Mauser, Wendy Osheroff, Jarrett

100 Henry Li et al.


Terry, Marcia Wilson-Heiner, and all SOP authorsin R&D for writing and reviewing standard operat-ing procedures. We also thank John Aldridge in theQA Document Group for formatting and process-ing the SOPs into the QA documentation database.

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