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US 2014/0272987 A1Published application
GLATIRAMER ACETATE RESPONSE BIOMARKER mRNA POTENCY ASSAY
AssigneeMylan Inc.
InventorsJeffrey P. Smith, Peter E. Lipsky
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Abstract
The invention describes assays for determining the potency of a test lot of glatiramer acetate (GA) by quantitating and comparing the levels of mRNA response biomarkers produced in mouse T-cells in response to stimulation with the test lot or a reference standard lot of GA, wherein the T-cells are obtained from mice immunized with the test lot or the reference standard lot of GA. Methods for identifying mRNA response biomarkers useful in the assays of the invention also are described.
Claims43
A process for determining the potency of a test lot of glatiramer acetate (GA), the process comprising:
- immunizing a test animal with a defined amount of a GA reference standard lot;
- culturing lymph node cells removed from the immunized test animal;
- incubating at least one sample containing a predetermined number of the cultured lymph node cells in the presence of a predetermined amount of the GA reference standard lot, and at least one sample containing a predetermined number of the cultured lymph node cells in the presence of a predetermined amount of a test lot of GA;
- measuring the amount of at least one GA response biomarker mRNA species in the lymph node cells incubated with the predetermined amount of the GA reference standard lot, and the amount of the least one GA response biomarker mRNA species in the lymph node cells incubated with the predetermined amount of the test lot of GA, and;
- comparing the amount of the least one GA response biomarker mRNA species measured in the lymph node cells incubated with the predetermined amount of the test lot of GA with the amount of the least one GA response biomarker mRNA species measured in the lymph node cells incubated with the predetermined amount of the GA reference standard lot;
- thereby determining the relative potency of the test lot of GA.
3 more elements
A process for preparing a pharmaceutical composition containing GA, wherein during the process a test lot of GA is tested to determine whether it has a desired potency relative to the potency of the GA reference standard lot, the process comprising:
- determining the potency of the test lot of GA and comparing it to the potency of the reference standard lot, wherein the potencies of the test lot of GA and the reference standard lot are determined by measuring the amount of at least one GA response biomarker mRNA species produced in a cell of a culture of lymph node cells removed from a test mammal that has been immunized with a defined amount of a GA reference standard lot, wherein at least one sample containing a predetermined number of the cultured lymph node cells is incubated in the presence of a predetermined amount of the GA reference standard lot, wherein at least one sample containing a substantially identical predetermined number of the cultured lymph node cells is incubated in the presence of the same predetermined amount of the test lot of GA;
- and
- admixing the test lot of GA into the pharmaceutical composition only if the test lot of GA is determined to have the desired potency relative to the potency of the reference standard lot.
A process for determining the potency and cross-potency of a test lot of GA, the process comprising:
- a) immunizing a first test animal with a defined amount of a GA reference standard lot;
- b) immunizing a second test animal with a defined amount of a GA test lot;
- c) culturing lymph node cells removed from the immunized first test animal, and separately culturing lymph node cells removed from the immunized second test animal;
- d) incubating at least one sample containing a predetermined number of the cultured lymph node cells from the immunized first test animal in the presence of a predetermined amount of the GA reference standard lot, and at least one sample containing a predetermined number of the cultured lymph node cells from the immunized first test animal in the presence of a predetermined amount of a test lot of GA;
- e) incubating at least one sample containing a predetermined number of the cultured lymph node cells from the immunized second test animal in the presence of a predetermined amount of the GA reference standard lot, and at least one sample containing a predetermined number of the cultured lymph node cells from the immunized second test animal in the presence of a predetermined amount of a test lot of GA;
- f) measuring the amount of (i) at least one GA response biomarker mRNA in the lymph node cells from the immunized first test animal incubated with the predetermined amount of the GA reference standard lot, and (ii) the same at least one GA response biomarker mRNA in the lymph node cells from the immunized first test animal incubated with the predetermined amount of the test lot of GA;
- g) measuring the amount of (i) the at least one GA response biomarker mRNA in the lymph node cells from the immunized second test animal incubated with the predetermined amount of the GA reference standard lot, and (ii) the at least one GA response biomarker mRNA in the lymph node cells from the immunized second test animal incubated with the predetermined amount of the test lot of GA;
- h) comparing the amount of the least one GA response biomarker mRNA measured in the lymph node cells from the immunized first test animal incubated with the predetermined amount of the test lot of GA with the amount of the least one GA response biomarker mRNA measured in the lymph node cells from the immunized first test animal incubated with the predetermined amount of the GA reference standard lot, thereby determining the relative potency of the test lot of GA;
- i) comparing the amount of the least one GA response biomarker mRNA measured in the lymph node cells from the immunized second test animal incubated with the predetermined amount of the test lot of GA with the amount of the least one GA response biomarker mRNA measured in the lymph node cells from the immunized second test animal incubated with the predetermined amount of the GA reference standard lot, thereby determining the relative potency of the reference standard lot of GA;
- j) comparing the amount of the least one GA response biomarker mRNA measured in the lymph node cells from the immunized first test animal incubated with the predetermined amount of the test lot of GA with the amount of the least one GA response biomarker mRNA measured in the lymph node cells from the immunized second test animal incubated with the predetermined amount of the GA test lot;
- thereby determining the cross-potency of the test lot of GA.
8 more elements
A process for determining the potency of a second lot of GA, the process comprising:
- immunizing a test animal with a defined amount of a first GA lot;
- culturing lymph node cells removed from the immunized test animal;
- incubating at least one sample containing a predetermined number of the cultured lymph node cells in the presence of a predetermined amount of the first GA lot, and at least one sample containing a predetermined number of the cultured lymph node cells in the presence of a predetermined amount of the second lot of GA;
- measuring the amount of at least one GA response biomarker mRNA in the lymph node cells incubated with the predetermined amount of the first GA lot, and the amount of the least one GA response biomarker mRNA in the lymph node cells incubated with the predetermined amount of the second lot of GA, and;
- comparing the amount of the least one GA response biomarker mRNA measured in the lymph node cells incubated with the predetermined amount of the second lot of GA with the amount of the least one GA response biomarker mRNA measured in the lymph node cells incubated with the predetermined amount of the first GA lot;
- thereby determining the relative potency of the second lot of GA.
3 more elements
Dependencies are inferred from the claim text.