Methods for donor cell analysis
OwnerMeng-Yin LIN, Hayung YOON +2inventors
InventorsMeng-Yin LIN, Hayung YOON, Thomas Charles PERTEL, Barbra Johnson SASU
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Abstract
Provided herein are methods, kits and reagents for analyzing the attributes of cell populations, such as donor cells prior to modification to provide engineered cells, e.g., engineered immune cells, such as CAR T cells. For example, provided herein are methods of determining the amount or percentage of biomarkers and/or secretion profiles of donor cell populations, selecting donor cells with certain biomarkers and/or secretion profiles, and engineering the CAR T cells from the selected donor cells.
Claims125
A method of manufacturing engineered immune cells comprising:
- a) detecting an HLA-DR expression level of 65% or less in an immune cell population;
- and
- b) modifying the immune cell population to express an exogenous nucleic acid sequence, thereby providing an engineered immune cell population.
An engineered immune cell population comprising 65% or less HLA-DR+ cells.
A method of manufacturing immune cells with improved in vitro functionality comprising:
- a) detecting a level of HLA-DR expression in an immune cell population to provide a detected level of HLA-DR expression;
- and
- b) modifying the immune cell population to express an exogenous nucleic acid sequence, thereby providing an engineered immune cell population,
- wherein the engineered immune cell population comprises or exhibits improved in vitro functionality as compared to an additional engineered immune cell population originated from an additional immune cell population having an additional level of HLA-DR expression that is higher than the detected level.
A method of selecting a donor immune cell population for engineering comprising:
- a) detecting a first level of HLA-DR expression in a first immune cell population to provide a first detected level of HLA-DR;
- b) detecting a second level of HLA-DR expression in a second immune cell population to provide a second detected level of HLA-DR, wherein the second detected level is greater than the first detected level;
- and
- c) selecting the first immune cell population for engineering.
A method of manufacturing immune cells with improved in vitro functionality comprising:
- a) modifying an immune cell population to express an exogenous nucleic acid sequence, thereby providing an engineered immune cell population;
- and
- b) depleting HLA-DR-positive engineered immune cells from the engineered immune cell population to provide an HLA-DR-depleted engineered immune cell population,
- wherein the HLA-DR-depleted engineered immune cell population comprises or exhibits improved in vitro functionality as compared to an engineered immune cell population that has not been depleted of HLA-DR-positive engineered immune cells.
A chimeric antigen receptor T (CAR-T) cell population, in which HLA-DR is expressed at a first level and the CAR-T cell population has improved in vitro functionality as compared to a CAR-T cell population in which HLA-DR is expressed at a second level, wherein the first level is lower than the second level.
A kit for in vitro functionality analysis of cell populations comprising:
- a) anti-HLA-DR binding agent;
- and
- b) instructions to use the binding agent to detect a level of HLA-DR expression in a cell population.
Dependencies are inferred from the claim text.