EUROPEAN PHARMACOPOEIA 11.

0 Meningococcal group C conjugate vaccine

limits approved by the competent authority for the particular The stability of the final lot and relevant intermediates is
product. Where justified and authorised, the test may be evaluated using 1 or more indicator tests. Such tests may
carried out on the monovalent bulk conjugate only. include determination of molecular size, determination of
Molecular-size or molecular-mass distribution. The free saccharide in the conjugate or an immunogenicity test
molecular-size or molecular-mass distribution is determined in animals.
by size-exclusion chromatography (2.2.30) combined with BACTERIAL SEED LOTS
an appropriate detection system. An acceptable value is The bacterial strains used for master seed lots shall be
established for each conjugate. Where justified and authorised, identified by historical records that include information on
the test may be carried out on the monovalent bulk conjugate their origin and the tests used to characterise the strain.
only. Cultures from the working seed lot shall have the same
Sterility (2.6.1). It complies with the test for sterility. characteristics as the strain that was used to prepare the
master seed lot.
ASSAY Purity of bacterial cultures is verified by methods of suitable
Saccharide. The total polysaccharide content for each group sensitivity. These may include inoculation into suitable media,
is determined by a suitable physico-chemical (for example examination of colony morphology, microscopic examination
liquid chromatography (2.2.29)) or immunochemical (for of Gram-stained smears and culture agglutination with
example enzyme-linked immunosorbent assay (ELISA) suitable specific antisera.
(2.7.1)) method. MENINGOCOCCAL GROUP C POLYSACCHARIDE
The content of each group is within the limits approved by the N. meningitidis is grown in a liquid medium that does not
competent authority for the particular product. contain high-molecular-mass polysaccharides and is free
from ingredients that will form a precipitate upon addition of
LABELLING cetyltrimethylammonium bromide (CTAB). The culture may
The label states : be inactivated by heat and filtered before the polysaccharide is
– the nominal amount of polysaccharide for each group (A, precipitated by addition of CTAB. The precipitate is further
C, W135 and Y) per single human dose ; purified using suitable methods to remove nucleic acids,
proteins and lipopolysaccharides and the final purification
– the type and amount of carrier protein per single human step consists of ethanol precipitation. An O-deacetylation step
dose. may also be included. Volatile matter, including water, in the
purified polysaccharide is determined by a suitable method
such as thermogravimetry (2.2.34). The value is used to
01/2019:2112 calculate the results of other tests with reference to the dried
substance, as prescribed below.
Only meningococcal group C polysaccharide that complies
with the following requirements may be used in the
preparation of the conjugate.
Protein (2.5.16) : maximum 1.0 per cent, calculated with
MENINGOCOCCAL GROUP C reference to the dried substance.
CONJUGATE VACCINE Nucleic acid (2.5.17) : maximum 1.0 per cent, calculated with
reference to the dried substance.
Vaccinum meningococcale classis C O-acetyl groups. Examine by a suitable method (for
coniugatum example 2.5.19). An acceptable value is established for the
particular product and each batch of meningococcal group C
DEFINITION polysaccharide must be shown to comply with this limit.
Meningococcal group C conjugate vaccine is a liquid or Sialic acid (2.5.23) : minimum 0.800 g of sialic acid per
freeze-dried preparation of purified capsular polysaccharide gram of meningococcal group C polysaccharide using
derived from a suitable strain of Neisseria meningitidis N-acetylneuraminic acid R to prepare the reference solution.
group C covalently linked to a carrier protein. Meningococcal
group C polysaccharide consists of partly O-acetylated Residual reagents. Where applicable, tests are carried out
or O-deacetylated repeating units of sialic acids, linked to determine residues of reagents used during inactivation
with 2α→9 glycosidic bonds. The carrier protein, when and purification. An acceptable value for each reagent is
conjugated to group C polysaccharide, is capable of established for the particular product and each batch of
inducing a T-cell-dependent B-cell immune response to the meningococcal group C polysaccharide must be shown
polysaccharide. The vaccine may contain an adjuvant. to comply with this limit. Where validation studies have
demonstrated removal of a residual reagent, the test on
PRODUCTION purified meningococcal group C polysaccharide may be
omitted.
GENERAL PROVISIONS
The production method shall consistently have been Molecular-size or molecular-mass distribution.
shown to yield meningococcal group C conjugate vaccines Molecular-size or molecular-mass distribution is determined
of satisfactory immunogenicity and safety in man. The by size-exclusion chromatography (2.2.30) combined with
production of meningococcal group C polysaccharide and of an appropriate detection system. Where applicable, the
the carrier protein are based on seed-lot systems. molecular-size distribution is also determined after chemical
modification of the polysaccharide. An acceptable value is
During development studies and wherever revalidation is
established for the meningococcal group C polysaccharide.
necessary, a test for pyrogens in rabbits (2.6.8) is carried out
Each batch must be shown to comply with this limit.
by injection of a suitable dose of the final lot. The vaccine is
shown to be acceptable with respect to absence of pyrogenic Identification and serological specificity. The identity
activity. and serological specificity are determined by a suitable
During development studies and wherever revalidation of the immunochemical method (2.7.1) or other suitable method, for
manufacturing process is necessary, it shall be demonstrated example 1H nuclear magnetic resonance spectrometry (2.2.33).
by tests in animals that the vaccine consistently induces a Bacterial endotoxins (2.6.14) : less than 100 IU per microgram
T-cell-dependent B-cell immune response. of meningococcal group C polysaccharide.

General Notices (1) apply to all monographs and other texts 1075
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Meningococcal polysaccharide vaccine EUROPEAN PHARMACOPOEIA 11.0

CARRIER PROTEIN FINAL LOT

The production and characteristics of the carrier proteins Only a final lot that is within the limits approved for the
are described in general chapter 5.2.11. Carrier proteins particular product and is satisfactory with respect to each of
for the production of conjugated polysaccharide vaccines for the requirements given below under Identification, Tests and
human use. Only a carrier protein that complies with the Assay may be released for use.
requirements of this chapter may be used in the preparation
of the conjugate. IDENTIFICATION
BULK CONJUGATE The vaccine is identified by a suitable immunochemical
Meningococcal group C polysaccharide is chemically modified method (2.7.1).
to enable conjugation ; it is usually partly depolymerised either TESTS
before or during this procedure. The conjugate is obtained
by the covalent binding of activated meningococcal group C pH (2.2.3). The pH of the vaccine, reconstituted if necessary,
oligosaccharide and the appropriate carrier protein. The is within the limits approved for the particular product.
conjugate purification procedures are designed to remove Aluminium (2.5.13) : maximum 1.25 mg per single human
residual reagents used for conjugation. The removal of dose, if aluminium hydroxide or hydrated aluminium
residual reagents and reaction by-products is confirmed by phosphate is used as the adsorbent.
suitable tests or by validation of the purification process. Water (2.5.12) : maximum 3.0 per cent for freeze-dried
Only a bulk conjugate that complies with the following vaccines.
requirements may be used in the preparation of the final bulk
vaccine. For each test and for each particular product, limits Free saccharide. Unbound saccharide is determined after
of acceptance are established and each batch of conjugate must removal of the conjugate, for example by anion-exchange
be shown to comply with these limits. liquid chromatography, size-exclusion or hydrophobic
chromatography, ultrafiltration or other validated methods.
Molecular-size or molecular-mass distribution. An acceptable value consistent with adequate immunogenicity,
Molecular-size or molecular-mass distribution is determined as shown in clinical trials, is established for the particular
by size-exclusion chromatography (2.2.30) combined with product and each final lot must be shown to comply with this
an appropriate detection system. An acceptable value is limit.
established for the bulk conjugate. Each batch must be shown
to comply with this limit. Sterility (2.6.1). It complies with the test for sterility.
Saccharide. The saccharide content is determined by a Bacterial endotoxins (2.6.14) : less than 25 IU per single
suitable validated assay (for example 2.5.23). Anion-exchange human dose.
liquid chromatography (2.2.29) with pulsed amperometric ASSAY
detection may also be used for determination of saccharide
content. An acceptable value is established for the particular Saccharide : minimum 80 per cent of the amount of
product and each batch of bulk conjugate must be shown to meningococcal group C polysaccharide stated on the label.
comply with this limit. The saccharide content is determined by a suitable validated
assay, for example sialic acid assay (2.5.23) or anion-exchange
Protein. The protein content is determined by a suitable liquid chromatography (2.2.29) with pulsed amperometric
chemical method (for example 2.5.16). An acceptable value is detection.
established for the particular product and each batch of bulk
conjugate must be shown to comply with this limit. LABELLING
Saccharide-to-protein ratio. Determine the ratio by The label states :
calculation. – the number of micrograms of meningococcal group C
Free saccharide. Unbound saccharide is determined after polysaccharide per single human dose ;
removal of the conjugate, for example by anion-exchange – the type and nominal amount of carrier protein per single
liquid chromatography, size-exclusion or hydrophobic human dose.
chromatography, ultrafiltration or other validated methods.
An acceptable value is established for the particular product
and each batch of bulk conjugate must be shown to comply 01/2019:0250
with this limit.
Free carrier protein. Determine the content, either directly
by a suitable method or by deriving the content by calculation
from the results of other tests. An acceptable value is
established for the particular product and each batch of bulk MENINGOCOCCAL
conjugate must be shown to comply with this limit. POLYSACCHARIDE VACCINE
Residual reagents. Removal of residual reagents such as
cyanide is confirmed by suitable tests or by validation of the Vaccinum meningococcale
process.
Sterility (2.6.1). It complies with the test for sterility, carried
polysaccharidicum
out using 10 mL for each medium or the equivalent of DEFINITION
100 doses, whichever is less. Meningococcal polysaccharide vaccine is a freeze-dried
FINAL BULK VACCINE preparation of one or more purified capsular polysaccharides
An adjuvant and a stabiliser may be added to the bulk obtained from one or more suitable strains of Neisseria
conjugate before dilution to the final concentration with a meningitidis group A, group C, group Y and group W135 that
suitable diluent. are capable of consistently producing polysaccharides.
Only a final bulk vaccine that complies with the following N. meningitidis group A polysaccharide consists of partly
requirement and is within the limits approved for the O-acetylated repeating units of N-acetylmannosamine, linked
particular product may be used in the preparation of the final with 1α→6 phosphodiester bonds.
lot. N. meningitidis group C polysaccharide consists of partly
Sterility (2.6.1). It complies with the test for sterility, carried O-acetylated repeating units of sialic acid, linked with 2α→9
out using 10 mL for each medium. glycosidic bonds.

1076 See the information section on general monographs (cover pages)

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