An improved assay method for determining the potency of recombinant protein
AssigneeKashiv Biosciences, LLC
InventorsShalini Sharma, Praveen Kumar Anidil Kizhakinagath, Kavya Siddhartha Jash
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Abstract
The present invention provides an improved assay method for determining the potency of recombinant proteins by culturing the immune cells such as antigen presenting cell and/or T cell in culture media comprising FBS less than 10%. Furthermore, the invention provides a method to eliminate or reduce the false estimation of potency of target recombinant protein. The assay method determines the potency of recombinant protein by evaluating the expression of inflammatory protein. More specifically the present invention describes the IL-2 inhibition assays for determination of potency of CTLA4 IgG1.
Claims21
An improved assay method for determining potency of recombinant protein comprising;
- a) culturing suitable antigen presenting cells and/or T cells in media comprising Fetal bovine serum less than 10%;
- b) incubating treated antigen presenting cells and/or T cells for suitable time and suitable temperature;
- c) mixing suitable recombinant protein with suitable stimulant to stimulate T cells in the cultured antigen presenting cells and/or T cells;
- d) incubating treated mixture for suitable time and suitable temperature;
- e) adding reagent in the incubated multi well plate;
- f) determining the inhibition of inflammatory protein;
- wherein cultured antigen presenting cells and/or T cells in Fetal bovine serum less than 10% provides improved potency compared to antigen presenting cells and/or T cells cultured in 10% Fetal bovine serum.
An improved assay method of recombinant protein comprising;
- the cultured antigen presenting cells and/or T cells in Fetal bovine serum less than 10% provides improved potency by improving 95% confidence intervals of inflammatory protein inhibition.
A process for reducing false potency estimation of recombinant protein comprising;
- a) culturing suitable antigen presenting cells and/or T cells in media comprising Fetal bovine serum less than 10%;
- b) incubating treated antigen presenting cells and/or T cells for suitable time and suitable temperature;
- c) mixing suitable recombinant protein with suitable stimulant to stimulate T cells in the cultured antigen presenting cells and/or T cells;
- d) incubating treated mixture for suitable time and suitable temperature;
- e) adding reagent in the incubated multi well plate;
- f) determining the inhibition of inflammatory protein;
- wherein cultured antigen presenting cells and/or T cells in Fetal bovine serum less than 10% provides improved potency compared to antigen presenting cells and/or T cells cultured in 10% Fetal bovine serum.
Dependencies are inferred from the claim text.