Secretions of proteins by encapsulated cells
AssigneeOne Cell Systems, Inc.
InventorsDiana Jesperson, Yevgenya Akselband, Patricia McGrath, Jan Trnovsky, Phillip T. Moen
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Abstract
The invention provides methods of analzying a secreted protein from a cell encapsulated in a microdrop. The microdrop is formulated with biotinylated matrix molecules at a reduced ratio of biotin to matrix molecules compared with previous formulations. The reduced ratio is advantageous for improving the resolution of detection and allows simultaneous detection of multiple secreted proteins and/or multiple cell surface markers. The invention further provides inter alia methods of isolating IgG isotype antibodies that have switched from IgM isotype.
Claims42
A method of analyzing a secreted molecule, comprising:
- encapsulating a cell in a microdrop wherein the microdrop comprises agarose molecules, first biotin molecules linked to the agarose molecules, capture molecules with affinity for a molecule secreted by the cell linked to second biotin molecules, and streptavidin linking the first and second biotin molecules, the first biotin molecules and the agarose molecules being in a molar ratio of less than 0.2 to 0.01 moles biotin per mole the agarose molecules, wherein the molecule is secreted from the cell and binds to the capture molecules thereby being retained within the microdrop;
- and
- detecting the secreted molecule.
A population of microdrops encapsulating cells, and the microdrops comprise matrix component molecules, first biotin molecules linked to the matrix component molecules, capture molecules with affinity for a protein secreted by the cell linked to second biotin molecules, and streptavidin linking the first and second biotin molecules, the first biotin molecules and the matrix molecules being in a molar ratio of 0.2 to 0.01 moles biotin per mole matrix component molecules.
In a method of analyzing a protein secreted by a cell in which the cell is encapsulated with a microdrop comprising biotinylated agarose the improvement wherein the molar ratio of biotin to agarose is 0.2 to 0.01 moles biotin per mole agarose.
Dependencies are inferred from the claim text.