Process for the measurement of the potency of glatiramer acetate

A process for preparing a pharmaceutical composition containing glatiramer acetate which comprises

• obtaining a batch of glatiramer acetate to be tested;
• immunizing female (SJLXBALB/c)F1 mice with a defined amount of a reference standard batch of glatiramer acetate,
• measuring separately the amount of interleukin-2 secreted by a primary culture of lymph node cells from the mice 9-11 days after immunization,
• i) where at least two samples containing a predetermined number of such cells are incubated separately in the presence of a predetermined amount of the reference standard batch, and
• ii) where at least two samples containing the substantially identical predetermined number of cells are incubated separately in the presence of the same predetermined amount of the batch of the glatiramer acetate being tested;
• comparing the amounts of interleukin-2 secreted which are so measured to determine the potency of the batch of glatiramer acetate being tested;
• and
• including in the pharmaceutical composition a batch only if its potency so measured is between 80% and 125% of the reference standard batch.

A process for determining the potency of a batch of glatiramer acetate, which comprises

• immunizing female (SJLXBALB/C)F1 mice with a defined amount of a reference standard batch of glatiramer acetate,
• measuring separately the amount of interleukin-2 secreted by a primary culture of lymph node cells from the mice 9-11 days after immunization,
• i) where at least two samples containing a predetermined number of such cells are incubated separately in the presence of a predetermined amount of the reference standard batch;
• and
• ii) where at least two samples containing the substantially identical predetermined number of cells are incubated separately in the presence of the same predetermined amount of the batch of glatiramer acetate;
• comparing the amounts of interleukin-2 secreted which are so measured, thereby determining the potency of the batch.

A process for determining whether a batch of glatiramer acetate, has a predetermined potency acceptable for inclusion in a pharmaceutical composition which comprises

• immunizing female (SJLXBALB/C)F1 mice with a defined amount of a reference standard batch of glatiramer acetate,
• measuring separately the amount of interleukin-2 secreted by a primary culture of lymph node cells from the mice 9-11 days after immunization,
• i) where at least two samples containing a predetermined number of such cells are incubated separately in the presence of a predetermined amount of the reference standard batch, and
• ii) where at least two samples containing the substantially identical predetermined number of cells are incubated separately in the presence of the same predetermined amount of the batch of glatiramer acetate;
• comparing the amounts of interleukin-2 secreted which are so measured to determine the potency of the batch;
• and
• comparing the potency so measured with the potency of the reference standard batch to determine whether the batch is acceptable for including in the pharmaceutical composition.

A process for preparing a pharmaceutical composition containing glatiramer acetate comprising obtaining a batch of glatiramer acetate;

• measuring the potency of the batch of glatiramer acetate relative to the potency of a reference standard of glatiramer acetate by:
• a) immunizing female (SJLXBALB/C)F1 mice between 8 and 12 weeks of age with a predetermined amount of the reference standard;
• b) preparing a primary culture of lymph node cells from the mice of step (a) 9-11 days after immunization;
• c) separately incubating at least five reference samples, each of which contains a predetermined number of cells from the primary culture of step (b) and a predetermined amount of the reference standard between 1 μg/ml and 25 μg/ml;
• d) incubating at least two samples, each of which contains a predetermined number of cells from the primary culture of step (b) and a predetermined amount of the glatiramer acetate;
• e) determining for each sample in steps (c) and (d), the amount of interleukin-2 secreted by the cells in each sample after 18-21 hours of incubation of such sample;
• and
• f) correlating the amounts of interleukin-2 secreted by the samples incubated with the glatiramer acetate with the amounts of interleukin-2 secreted by the samples incubated with the reference standard so as to determine the potency of the batch of glatiramer acetate relative to the reference standard,
• wherein in each sample in steps (c) and (d) the predetermined number of cells is substantially identical, and wherein for each sample containing a predetermined amount of glatiramer acetate there is a corresponding reference sample containing a substantially identical predetermined amount of the reference standard;
• and
• qualifying the batch of glatiramer acetate as acceptable for use in the pharmaceutical composition if its relative potency so measured is between 80% and 125% of the potency of the reference standard.