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US 8,465,937 B2Granted
Methods for identifying B-clones which bind cell-surface antigens
AssigneeESBATech, an Alcon Biomedical Research Unit LLC
InventorsDavid Urech, Valerie Hulmann-Cottier
Life of this patent
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Abstract
The invention provides methods for identifying immunobinders, such as scFv antibodies, capable of specifically binding to cell surface antigens, and compositions identified according to said methods.
Claims18
A method for identifying a B-cell clone that specifically binds to an antigen of interest comprising:
- (a) providing a plurality of B-cells;
- (b) contacting the plurality of B-cells with the antigen of interest, wherein the antigen of interest comprises a first sortable label, and staining the plurality of B-cells with anti-IgG antibodies comprising a second sortable label and anti-IgM antibodies comprising a third sortable label;
- (c) separating from the plurality of B-cells one or more B-cells that can specifically bind to the antigen of interest using a cell sorter, wherein the presence of the first and second sortable label, but not the third sortable label, in a complex is indicative of the binding of a B-cell to an antigen of interest, thereby identifying a B-cell clone that binds to the antigen of interest.
A method for identifying a B-cell clone that specifically binds to an antigen of interest comprising:
- (a) immunizing an animal with an antigen of interest;
- (b) isolating a plurality of B-cells from the immunized animal;
- (c) contacting the plurality of B-cells with the antigen of interest, wherein the antigen of interest comprises a first sortable label, and staining the plurality of B-cells with anti-IgG antibodies comprising a second sortable label and anti-IgM antibodies comprising a third sortable label;
- (d) separating from the plurality of B-cells one or more B-cells that can specifically bind to the antigen of interest using a cell sorter, wherein the presence of the first and second sortable label, but not the third sortable label, in a complex is indicative of the binding of a B-cell to an antigen of interest, thereby identifying a B-cell clone that binds to the antigen of interest.
1 more element
Dependencies are inferred from the claim text.