`
`European Medicines Agency
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`
`July 1996
`CPMP/ICH/138/95
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`ICH Topic Q 5 C
`Quality of Biotechnological Products:
`Stability Testing of Biotechnological/Biological Products
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`NOTE FOR GUIDANCE ON QUALITY OF BIOTECHNOLO-GICAL PRODUCTS:
`STABILITY TESTING OF BIOTECHNOLOGICAL/BIOLOGICAL PRODUCTS
`(CPMP/ICH/138/95)
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`Step 5
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`TRANSMISSION TO CPMP
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`FINAL APPROVAL BY CPMP
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`PROPOSED DATE FOR COMING INTO OPERATION
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`December 1995
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`December 1995
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`July 1996
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`7 Westferry Circus, Canary Wharf, London, E14 4HB, UK
`Tel. (44-20) 74 18 85 75 Fax (44-20) 75 23 70 40
`E-mail: mail@emea.eu.int http://www.emea.eu.int
`EMEA 2006 Reproduc ion and/or distribution of this document is authorised for non commercial purposes only provided the EMEA is acknowledged
`
`Novartis Exhibit 2230.001
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`QUALITY OF BIOTECHNOLOGICAL PRODUCTS:
`STABILITY TESTING OF BIOTECHNOLOGICAL/BIOLOGICAL PRODUCTS
`Annex to the ICH Harmonised Tripartite Guideline for the Stability Testing of New Drug
`Substances and Products
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`
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`PREAMBLE
`1.
`The guidance stated in the ICH Harmonised Tripartite Guideline ‘Stability Testing of New
`Drug Substances
`and Products’
`(27 October 1993)
`applies
`in general
`to
`biotechnological/biological products. However, biotechnological/biological products do have
`distinguishing characteristics to which consideration should be given in any well-defined
`testing program designed to confirm their stability during the intended storage period. For
`such products, in which the active components are typically proteins and/or polypeptides,
`maintenance of molecular conformation and, hence of biological activity, is dependent on
`noncovalent as well as covalent forces. The products are particularly sensitive to
`environmental factors such as temperature changes, oxidation, light, ionic content, and shear.
`In order to ensure maintenance of biological activity and to avoid degradation, stringent
`conditions for their storage are usually necessary.
`
`The evaluation of stability may necessitate complex analytical methodologies. Assays for
`biological activity, where applicable, should be part of the pivotal stability studies.
`Appropriate physicochemical, biochemical and immunochemical methods for the analysis of
`the molecular entity and the quantitative detection of degradation products should also be part
`of the stability program whenever purity and molecular characteristics of the product permit
`use of these methodologies.
`
`With the above concerns in mind, the applicant should develop the proper supporting stability
`data for a biotechnological/biological product and consider many external conditions which
`can affect the product's potency, purity and quality. Primary data to support a requested
`storage period for either drug substance or drug product should always be based on long-term,
`real-time, real-condition stability studies. Thus, the development of a proper long-term
`stability program becomes critical to the successful development of a commercial product.
`The purpose of this document is to give guidance to applicants regarding the type of stability
`studies that should be provided in support of marketing applications. It is understood that
`during the review and evaluation process, continuing updates of initial stability data may
`occur.
`
`SCOPE OF THE ANNEX
`2.
`The guidance stated in this annex applies to well-characterised proteins and polypeptides,
`their derivatives and products of which they are components, and which are isolated from
`tissues, body fluids, cell cultures, or produced using rDNA technology. Thus, the document
`covers the generation and submission of stability data for products such as cytokines
`(interferons,
`interleukins,
`colony-stimulating
`factors,
`tumour
`necrosis
`factors),
`erythropoietins, plasminogen activators, blood plasma factors, growth hormones and growth
`factors, insulins, monoclonal antibodies, and vaccines consisting of well-characterised
`proteins or polypeptides. In addition, the guidance outlined in the following sections may
`apply to other types of products, such as conventional vaccines, after consultation with the
`appropriate regulatory authorities. The document does not cover antibiotics, allergenic
`extracts, heparins, vitamins or whole blood.
`
`© EMEA 2006
`
`
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`2
`
`Novartis Exhibit 2230.002
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`
`TERMINOLOGY
`3.
`For the basic terms used in this annex the reader is referred to the "Glossary" in the ICH
`Harmonised Tripartite Guideline ‘Stability Testing of New Drug Substances and Products’
`(27 October 1993). However, since manufacturers of biotechnological/biological products
`sometimes use traditional terminology, traditional terms are specified in parentheses to assist
`the reader. A supplemental glossary is also included that explains certain terms used in the
`production of biotechnological/biological products.
`
`4.
`
`SELECTION OF BATCHES
`
`4.1 Drug Substance (Bulk Material)
`Where bulk material is to be stored after manufacture but prior to formulation and final
`manufacturing, stability data should be provided on at least three batches for which
`manufacture and storage are representative of the manufacturing scale of production. A
`minimum of six months stability data at the time of submission should be submitted in cases
`where storage periods greater than six months are requested. For drug substances with storage
`periods of less than six months, the minimum amount of stability data in the initial submission
`should be determined on a case-by-case basis. Data from pilot-plant-scale batches of drug
`substance produced at a reduced scale of fermentation and purification may be provided at the
`time the dossier is submitted to the regulatory agencies with a commitment to place the first
`three manufacturing scale batches into the long-term stability program after approval.
`
`The quality of the batches of drug substance placed into the stability program should be
`representative of the quality of the material used in pre-clinical and clinical studies and of the
`quality of the material to be made at manufacturing scale. In addition, the drug substance
`(bulk material) made at pilot-plant scale should be produced by a process and stored under
`conditions representative of that used for the manufacturing scale. The drug substance entered
`into the stability program should be stored in containers which properly represent the actual
`holding containers used during manufacture. Containers of reduced size may be acceptable for
`drug substance stability testing provided that they are constructed of the same material and
`use the same type of container/closure system that is intended to be used during manufacture.
`
`Intermediates
`4.2
`During manufacture of biotechnological/biological products, the quality and control of certain
`intermediates may be critical to the production of the final product. In general, the
`manufacturer should identify intermediates and generate in-house data and process limits that
`assure their stability within the bounds of the developed process. While the use of pilot-plant-
`scale data is permissible, the manufacturer should establish the suitability of such data using
`the manufacturing-scale process.
`
`4.3 Drug Product (Final Container Product)
`Stability information should be provided on at least three batches of final container product
`representative of that which will be used at manufacturing scale. Where possible, batches of
`final container product included in stability testing should be derived from different batches of
`bulk material. A minimum of six months data at the time of submission should be submitted
`in cases where storage periods greater than six months are requested. For drug products with
`storage periods of less than six months, the minimum amount of stability data in the initial
`submission should be determined on a case-by-case basis. Product expiration dating will be
`based upon the actual data submitted in support of the application. Since dating is based upon
`the real-time/real-temperature data submitted for review, continuing updates of initial stability
`data should occur during the review and evaluation process. The quality of the final container
`
`© EMEA 2006
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`3
`
`Novartis Exhibit 2230.003
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`
`product placed on stability studies should be representative of the quality of the material used
`in the preclinical and clinical studies. Data from pilot-plant scale batches of drug product may
`be provided at the time the dossier is submitted to the regulatory agencies with a commitment
`to place the first three manufacturing scale batches into the long-term stability program after
`approval. Where pilot-plant scale batches were submitted to establish the dating for a product
`and, in the event that product produced at manufacturing scale does not meet those long-term
`stability specifications throughout the dating period or is not representative of the material
`used in pre-clinical and clinical studies, the applicant should notify the appropriate regulatory
`authorities to determine a suitable course of action.
`
`4.4 Sample selection criteria
`Where one product is distributed in batches differing in fill volume (e.g., 1 milliliter (ml), 2
`ml, or 10 ml), unitage (e.g., 10 units, 20 units, or 50 units), or mass (e.g., 1 milligram (mg), 2
`mg, or 5 mg) samples to be entered into the stability program may be selected on the basis of
`a matrix system and/or by bracketing.
`
`Matrixing, i.e., the statistical design of a stability study in which different fractions of samples
`are tested at different sampling points, should only be applied when appropriate
`documentation is provided that confirms that the stability of the samples tested represents the
`stability of all samples. The differences in the samples for the same drug product should be
`identified as, for example, covering different batches, different strengths, different sizes of the
`same closure and possibly, in some cases, different container/closure systems. Matrixing
`should not be applied to samples with differences that may affect stability, such as different
`strengths and different containers/closures, where it cannot be confirmed that the products
`respond similarly under storage conditions.
`
`Where the same strength and exact container/closure system is used for three or more fill
`contents, the manufacturer may elect to place only the smallest and largest container size into
`the stability program, i.e., bracketing. The design of a protocol that incorporates bracketing
`assumes that the stability of the intermediate condition samples are represented by those at the
`extremes. In certain cases, data may be needed to demonstrate that all samples are properly
`represented by data collected for the extremes.
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`STABILITY-INDICATING PROFILE
`5.
`On the whole, there is no single stability-indicating assay or parameter that profiles the
`stability characteristics of a biotechnological/biological product. Consequently,
`the
`manufacturer should propose a stability-indicating profile that provides assurance that
`changes in the identity, purity and potency of the product will be detected.
`
`At the time of submission, applicants should have validated the methods that comprise the
`stability-indicating profile and the data should be available for review. The determination of
`which tests should be included will be product-specific. The items emphasised in the
`following subsections are not
`intended
`to be all-inclusive, but represent product
`characteristics that should typically be documented to adequately demonstrate product
`stability.
`
`© EMEA 2006
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`4
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`Novartis Exhibit 2230.004
`Regeneron v. Novartis, IPR2021-00816
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`

`

`
`
`5.1 Protocol
`The dossier accompanying the application for marketing authorisation should include a
`detailed protocol for the assessment of the stability of both drug substance and drug product
`in support of the proposed storage conditions and expiration dating periods. The protocol
`should
`include all necessary
`information which demonstrates
`the stability of
`the
`biotechnological/biological product throughout the proposed expiration dating period
`including, for example, well-defined specifications and test intervals. The statistical methods
`that should be used are described in the Tripartite Guideline on stability.
`
`5.2 Potency
`When the intended use of a product is linked to a definable and measurable biological
`activity, testing for potency should be part of the stability studies. For the purpose of stability
`testing of the products described in this guideline, potency is the specific ability or capacity of
`a product to achieve its intended effect. It is based on the measurement of some attribute of
`the product and is determined by a suitable quantitative method. In general, potencies of
`biotechnological/biological products tested by different laboratories can be compared in a
`meaningful way only if expressed in relation to that of an appropriate reference material. For
`that purpose, a reference material calibrated directly or indirectly against the corresponding
`national or international reference material should be included in the assay.
`
`Potency studies should be performed at appropriate intervals as defined in the stability
`protocol and the results should be reported in units of biological activity calibrated, whenever
`possible, against nationally or internationally recognised standard. Where no national or
`international standards exists, the assay results may be reported in in-house derived units
`using a characterised reference material.
`
`In some biotechnological/biological products, potency is dependent upon the conjugation of
`the active ingredient(s) to a second moiety or binding to an adjuvant. Dissociation of the
`active ingredient(s) from the carrier used in conjugates or adjuvants should be examined in
`real-time/real-temperature studies (including conditions encountered during shipment). The
`assessment of the stability of such products may be difficult since, in some cases, in vitro tests
`for biological activity and physicochemical characterisation are impractical or provide
`inaccurate
`results. Appropriate
`strategies
`(e.g.,
`testing
`the product prior
`to
`conjugation/binding, assessing the release of the active compound from the second moiety, in
`vivo assays) or the use of an appropriate surrogate test should be considered to overcome the
`inadequacies of in vitro testing.
`
`5.3 Purity and Molecular Characterisation
`For the purpose of stability testing of the products described in this guideline, purity is a
`relative term. Due to the effect of glycosylation, deamidation, or other heterogeneities, the
`absolute purity of a biotechnological/biological product is extremely difficult to determine.
`Thus, the purity of a biotechnological/biological product should be typically assessed by more
`than one method and the purity value derived is method-dependent. For the purpose of
`stability testing, tests for purity should focus on methods for determination of degradation
`products.
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`The degree of purity, as well as individual and total amounts of degradation products of the
`biotechnological/biological product entered into the stability studies, should be reported and
`documented whenever possible. Limits of acceptable degradation should be derived from the
`analytical profiles of batches of the drug substance and drug product used in the pre-clinical
`and clinical studies.
`
`© EMEA 2006
`
`
`
`5
`
`Novartis Exhibit 2230.005
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`
`immunochemical analytical
`The use of relevant physicochemical, biochemical and
`methodologies should permit a comprehensive characterisation of the drug substance and/or
`drug product (e.g., molecular size, charge, hydrophobicity) and the accurate detection of
`degradation changes that may result from deamidation, oxidation, sulfoxidation, aggregation
`or fragmentation during storage. As examples, methods that may contribute to this include
`electrophoresis (SDS-PAGE, immunoelectrophoresis, Western blot, isoelectrofocusing), high-
`resolution chromatography (e.g., reversed-phase chromatography, gel filtration, ion exchange,
`affinity chromatography), and peptide mapping.
`
`Wherever significant qualitative or quantitative changes indicative of degradation product
`formation are detected during long-term, accelerated and/or stress stability studies,
`consideration should be given to potential hazards and to the need for characterisation and
`quantification of degradation products within the long-term stability program. Acceptable
`limits should be proposed and justified, taking into account the levels observed in material
`used in pre-clinical and clinical studies.
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`For substances that can not be properly characterised or products for which an exact analysis
`of the purity cannot be meaningfully determined through routine analytical methods, the
`applicant should propose and justify alternative testing procedures.
`
`5.4 Other Product Characteristics
`to
`relating
`specifically
`not
`though
`The
`following
`product
`characteristics,
`biotechnological/biological products, should be monitored and reported for the drug product
`in its final container:
`• Visual appearance of the product (colour and opacity for solutions/suspensions; colour,
`texture and dissolution time for powders), visible particulates in solutions or after the
`reconstitution of powders or lyophilised cakes, pH, and moisture level of powders and
`lyophilised products.
`• Sterility testing or alternatives (e.g., container/closure integrity testing) should be
`performed at a minimum initially and at the end of the proposed shelf-life.
`• Additives (e.g., stabilisers, preservatives) or excipients may degrade during the dating
`period of the drug product. If there is any indication during preliminary stability studies
`that reaction or degradation of such materials adversely affect the quality of the drug
`product, these items may need to be monitored during the stability program.
`• The container/closure has the potential to adversely affect the product and should be
`carefully evaluated (see below).
`
`6.
`
`STORAGE CONDITIONS
`
`6.1 Temperature
`Since most finished biotechnological/biological products need precisely defined storage
`temperatures, the storage conditions for the real-time/real-temperature stability studies may be
`confined to the proposed storage temperature.
`
`6.2 Humidity
`Biotechnological/biological products are generally distributed in containers protecting them
`against humidity. Therefore, where it can be demonstrated that the proposed containers (and
`
`© EMEA 2006
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`6
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`Novartis Exhibit 2230.006
`Regeneron v. Novartis, IPR2021-00816
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`

`

`
`conditions of storage) afford sufficient protection against high and low humidity, stability
`tests at different relative humidities can usually be omitted. Where humidity-protecting
`containers are not used, appropriate stability data should be provided.
`
`6.3 Accelerated and stress conditions
`As previously noted, the expiration dating should be based on real-time/real-temperature data.
`However, it is strongly suggested that studies be conducted on the drug substance and drug
`product under accelerated and stress conditions. Studies under accelerated conditions may
`provide useful support data for establishing the expiration date, provide product stability
`information for future product development (e.g., preliminary assessment of proposed
`manufacturing changes such as change in formulation, scale-up), assist in validation of
`analytical methods for the stability program, or generate information which may help
`elucidate the degradation profile of the drug substance or drug product. Studies under stress
`conditions may be useful in determining whether accidental exposures to conditions other
`than those proposed (e.g., during transportation) are deleterious to the product and also for
`evaluating which specific test parameters may be the best indicators of product stability.
`Studies of the exposure of the drug substance or drug product to extreme conditions may help
`to reveal patterns of degradation; if so, such changes should be monitored under proposed
`storage conditions. While the Tripartite Guideline on stability describes the conditions of the
`accelerated and stress study, the applicant should note that those conditions may not be
`appropriate for biotechnological/biological products. Conditions should be carefully selected
`on a case-by-case basis.
`
`6.4 Light
`Applicants should consult the appropriate regulatory authorities on a case-by-case basis to
`determine guidance for testing.
`
`6.5 Container/Closure
`Changes in the quality of the product may occur due to the interactions between the
`formulated biotechnological/biological product and container/closure. Where the lack of
`interactions cannot be excluded in liquid products (other than sealed ampoules), stability
`studies should include samples maintained in the inverted or horizontal position (i.e., in
`contact with the closure), as well as in the upright position, to determine the effects of the
`closure on product quality. Data should be supplied for all different container/closure
`combinations that will be marketed.
`
`In addition to the standard data necessary for a conventional single-use vial, the applicant
`should demonstrate that the closure used with a multiple-dose vial is capable of withstanding
`the conditions of repeated insertions and withdrawals so that the product retains its full
`potency, purity, and quality for the maximum period specified in the instructions-for-use on
`containers, packages, and/or package inserts. Such labelling should be in accordance with
`relevant national/regional requirements.
`
`6.6 Stability after Reconstitution of Freeze-Dried Product
`The stability of freeze-dried products after their reconstitution should be demonstrated for the
`conditions and the maximum storage period specified on containers, packages, and/or package
`inserts. Such labelling should be in accordance with relevant national/regional requirements.
`
`
`
`© EMEA 2006
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`
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`7
`
`Novartis Exhibit 2230.007
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`
`TESTING FREQUENCY
`7.
`The shelf-lives of biotechnological/biological products may vary from days to several years.
`Thus, it is difficult to draft uniform guidelines regarding the stability study duration and
`testing frequency that would be applicable to all types of biotechnological/biological
`products. With only a few exceptions, however, the shelf-lives for existing products and
`potential future products will be within the range of 0.5 to five years. Therefore, the guidance
`is based upon expected shelf-lives in that range. This takes into account the fact that
`degradation of biotechnological/biological products may not be governed by the same factors
`during different intervals of a long storage period.
`
`When shelf-lives of one year or less are proposed, the real-time stability studies should be
`conducted monthly for the first three months and at three-month intervals thereafter.
`
`For products with proposed shelf-lives of greater than one year, the studies should be
`conducted every three months during the first year of storage, every six months during the
`second year, and annually thereafter.
`
`While the testing intervals listed above may be appropriate in the pre-approval or pre-license
`stage, reduced testing may be appropriate after approval or licensure where data are available
`that demonstrate adequate stability. Where data exist that indicate the stability of a product is
`not compromised, the applicant is encouraged to submit a protocol which supports
`elimination of specific test intervals (e.g., nine-month testing) for post-approval/post-
`licensure, long-term studies.
`
`SPECIFICATIONS
`8.
`Although biotechnological/biological products may be subject to significant losses of activity,
`physicochemical changes, or degradation during storage,
`international and national
`regulations have provided little guidance with respect to distinct release and end of shelf-life
`specifications. Recommendations for maximum acceptable losses of activity, limits for
`physicochemical changes, or degradation during the proposed shelf-life have not been
`developed for individual types or groups of biotechnological/biological products but are
`considered on a case-by-case basis. Each product should retain its specifications within
`established limits for safety, purity, and potency throughout its proposed shelf-life. These
`specifications and limits should be derived from all available information using the
`appropriate statistical methods. The use of different specifications for release and expiration
`should be supported by sufficient data to demonstrate that clinical performance is not affected
`as discussed in the Tripartite Guideline on stability.
`
`LABELLING
`9.
`For most biotechnological/biological drug substances and drug products, precisely defined
`storage temperatures are recommended. Specific recommendations should be stated,
`particularly for drug substances and drug products that cannot tolerate freezing. These
`conditions, and where appropriate, recommendations for protection against light and/or
`humidity, should appear on containers, packages, and/or package inserts. Such labelling
`should be in accordance with relevant national regional requirements.
`
`© EMEA 2006
`
`
`
`8
`
`Novartis Exhibit 2230.008
`Regeneron v. Novartis, IPR2021-00816
`
`

`

`
`
`GLOSSARY
`
`Conjugated Product
`A conjugated product is made up of an active ingredient (for example, peptide, carbohydrate)
`bound covalently or noncovalently to a carrier (for example, protein, peptide, inorganic
`mineral) with the objective of improving the efficacy or stability of the product.
`
`Degradation Product
`A molecule resulting from a change in the drug substance (bulk material) brought about over
`time. For the purpose of stability testing of the products described in this guideline, such
`changes could occur as a result of processing or storage (e.g., by deamidation, oxidation,
`aggregation, proteolysis). For biotechnological/biological products some degradation products
`may be active.
`
`Impurity
`Any component of the drug substance (bulk material) or drug product (final container
`product) which is not the chemical entity defined as the drug substance, an excipient, or other
`additives to the drug product.
`
`Intermediate
`For biotechnological/biological products, a material produced during a manufacturing process
`which is not the drug substance or the drug product but whose manufacture is critical to the
`successful production of the drug substance or the drug product. Generally, an intermediate
`will be quantifiable and specifications will be established to determine the successful
`completion of the manufacturing step prior to continuation of the manufacturing process. This
`includes material which may undergo further molecular modification or be held for an
`extended period of time prior to further processing.
`
`Manufacturing-Scale Production
`Manufacture at the scale typically encountered in a facility intended for product production
`for marketing.
`
`Pilot-Plant Scale
`The production of the drug substance or drug product by a procedure fully representative of
`and simulating that to be applied at manufacturing scale. The methods of cell expansion,
`harvest, and product purification should be identical except for the scale of production.
`
`© EMEA 2006
`
`
`
`9
`
`Novartis Exhibit 2230.009
`Regeneron v. Novartis, IPR2021-00816
`
`