ICH Q5C

Q5C
• QUALITY OF BIOTECHNOLOGICAL
PRODUCTS: STABILITY TESTING OF
BIOTECHNOLOGICAL/ BIOLOGICAL
PRODUCTS
• Biologicals, is pharmaceutical products obtained directly
from living organisms.

• Examples of biologicals are blood plasma products, vaccines,

antivenoms, immunoglobulins, and allergenic extracts.

• The key aspect of biological drug substances and products is

that they are more labile compared to most traditional small
molecule pharmaceuticals.

• Generally, they require low-temperature storage conditions

such as refrigeration (2◦−8◦C), freezing (−10◦ to −20◦C), or
even ultra-low (−40◦ to −80◦C) storage temperatures
Comparison of biologic products
vs. small molecule products
Degradation of Peptide and Protein Pharmaceuticals

Deamidation, racemization,
isomerization, hydrolysis,
Chemical disulfide
formation/exchange, β -
elimination, and oxidation

Degradation

Denaturation, aggregation,
Physical adsorption, and
precipitation

• It involves changes in covalent bonds,

and the latter involve changes in
noncovalent interactions such as
hydrophobic bonding/associations.
 SELECTION OF BATCHES

A1. Active substance (Bulk Material)

• 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 active 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
active 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 active substance placed into the
stability pre-clinical and clinical studies program should be
representative of the quality of the material used in and of the
quality of the material to be made at manufacturing scale.

• In addition, the active 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 active 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 active

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
A2. Intermediates

• 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.
A3. Medicinal 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 medicinal 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 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 medicinal 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.
A4. Sample selection criteria
Bracketing is a design of a stability testing schedule in which the
experimental stability data are obtained only from the
extremes of packaging size or strengths.
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 medicinal 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.
B. STABILITY-INDICATING PROFILE
• To check the identity, purity and potency of the
product

• 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.
 B1 Protocol
• The dossier accompanying the application for
marketing authorization should include a
detailed protocol for the assessment of the
stability of both active substance and
medicinal 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.
 B2 Potency
• Testing for potency should be part of the stability
studies.
• 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.
 B3 Purity and Molecular Characterisation
• Due to the effect of glycosylation, deamidation, or other heterogeneities,
the absolute purity of a biotechnological/biological product is extremely
difficult to determine.
• 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.
• The use of relevant physico-chemical, biochemical and immunochemical
analytical methodologies should permit a comprehensive characterisation
of the active substance and/or medicinal product (e.g. molecular size,
charge, hydrophobicity) and the accurate detection of degradation changes
that may result from deamidation, oxidation, sulphoxidation, 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
B4 Other Product Characteristics
The following product characteristics, though not specifically
relating to biotechnological/biological products, should be
monitored and reported for the medicinal 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 medicinal product.

•If there is any indication during preliminary

stability studies that reaction or degradation of
such materials adversely affect the quality of
the medicinal 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.
 C. STORAGE CONDITIONS
• Temperature
• Humidity
• Accelerated and stress conditions
• Light
• Container/Closure
• Stability after Reconstitution of Freeze-Dried
Product
 TESTING FREQUENCY
• Shelf-lives of biotechnological/biological products
may vary from days to several years.
• 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.
• 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
 LABELLING
• Recommendations for protection against light
and/or humidity,
• should appear on containers, packages,
and/or package inserts.