Methods for assessing cell surface glycosylation
AssigneeJuno Therapeutics, Inc.
InventorsPaul Ken KODAMA, Tom KOWSKI, Mirna MUJACIC, Kenneth Mayo PRENTICE
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Abstract
Provided herein are methods for assessing cell surface glycans, e.g., N-glycans, by assessing a sample of released surface glycans, and determining the presence, absence, or level of glycans present in the sample. Also provided are methods of assaying and/or evaluating a cell composition by assessing the cell surface glycan profile of the cell composition and comparing the profile to a reference sample. Methods for manufacturing and/or culturing a plurality of cell compositions having consistent surface glycan expression with low variability are also provided.
Claims191
A method for assessing cell surface glycans, the method comprising:
- (a) incubating a test composition comprising a plurality of cells under conditions to release one or more glycans from the surface of cells in the test composition, wherein a sample comprising one or more cell surface glycans is generated;
- and
- (b) determining the presence, absence, identity and/or level of glycans present in the sample, thereby assessing the cell surface glycan profile of the sample.
A method for assessing cell surface glycans, the method comprising determining the presence, absence, identity and/or level of glycans present in a sample, thereby assessing the cell surface glycan profile of the sample, wherein the sample comprises one or more glycans released from the surface of cells present in a test composition comprising a plurality of cells after incubation of the test composition under conditions to release the one or more glycans.
The method of any of claims 1-35, wherein the incubating the test composition is at a temperature of about 37° C.
A method for manufacturing a cell composition, comprising
- incubating and/or contacting an input composition comprising a plurality of cells with one or more agents and/or under one or more conditions thereby generating the cell composition,
- wherein the cell composition comprises one or a plurality of cells that are genetically, phenotypically, and/or functionally different from one or a plurality of cells from the input composition, and
- wherein the cell composition comprises one or a plurality of cells that comprise a cell surface glycan profile comprising one or more target glycans and/or each of the one or more target glycans in the cell surface glycan profile differs by no more than 25% from the cell surface glycan profile or each of the one or more target glycans to the total glycans present in a reference sample, wherein the cell surface glycan profile comprises glycans released from the surface of cells in the cell composition.
A method for screening one or more test agents or conditions on a cell composition, comprising:
- (a) assessing a cell surface glycan profile in a sample from a test cell composition, wherein the test cell composition is or is derived from a source composition that has been incubated or treated in the presence of one or more test agents or conditions;
- and
- (b) comparing the cell surface glycan profile of the sample to the cell surface glycan profile of a reference sample, the reference sample comprising one or more target glycans.
A method for screening one or more test agents or conditions on a cell composition, comprising comparing the cell surface glycan profile of a sample compared to the cell surface glycan profile of a reference sample, wherein the sample is from a test cell composition that is or is derived from a source composition that has been incubated or treated in the presence of one or more test agents or conditions.
Dependencies are inferred from the claim text.