Kits and method of modifying, detecting, and sorting antibody-producing cells
OwnerJunyu LINinventor
Life of this patent
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Abstract
Described herein are methods and compositions for modifying a mammalian cell plasma membrane to retain secreted antibodies, capturing antibodies produced by the cells and sorting antibody producing cells according to antibody specificities or antibody quantities. These methods and compositions may be particularly useful for biological and medical applications. The methods may include modifying cell membranes from a population of antibody producing cells by inserting a membrane anchor into the cell membranes, a generic antibody capture molecule attached to the cell anchor, capturing antibodies produced by the cell, and detecting and dispending cells according to antibody specificities or antibody quantities with a microparticle sorting and dispensing apparatus.
Claims23
A method of isolating an antibody producing cell, the method comprising:
- contacting a population of antibody-producing cells with a soluble presentation construct comprising a lipid modified protein A molecule (MPA), lipid modified protein G molecule (MPG) or a lipid modified protein A/G molecule (MPAG) so that the MPA, MPG or MPAG is presented on the surface of the antibody-producing cells;
- incubating the antibody-producing cells so that antibody produced by the cells is captured by the MPA, MPG or MPAG on the surface of the antibody-producing cells;
- exposing the antibody-producing cells to a detection agent comprising a detectably labeled antigen, a detectably labeled protein A molecule, a detectably labeled protein G molecule and/or a detectably labeled protein A/G molecule;
- sorting the cells using a cell sorting device according to the level of the detection agent.
A method of isolating an antibody producing cell, the method comprising:
- contacting a population of antibody-producing cells with a soluble presentation construct comprising a lipid modified protein A/G molecule (MPAG) having at least two oleic acid based chains coupled to a poly(ethylene glycol) (PEG) polymer that is coupled to the protein A/G molecule, so that the MPAG is presented on the surface of the antibody-producing cells;
- incubating the antibody-producing cells so that antibody produced by a cell in the population of antibody-producing cells is captured by the MPAG on the surface of that cell;
- exposing the antibody-producing cells to a detection agent detection agent comprising a detectably labeled marker that binds to the antibody produced by the cells;
- sorting the cells using a cell sorting device according to the level of the detection agent.
A kit for isolating antibody-producing cells, the kit comprising:
- a water-soluble presentation agent comprising a solution of a lipid modified protein A/G molecule (MPAG) having at least two oleic acid based chains coupled to a poly(ethylene glycol) (PEG) polymer that is coupled to the protein A/G molecule;
- and
- a detection agent comprising a solution of fluorescently labeled protein A/G.
A kit for isolating antibody-producing cells, the kit comprising:
- a water-soluble presentation agent comprising a solution of a lipid modified protein A/G molecule (MPAG) having at least two oleic acid based chains coupled to a poly(ethylene glycol) (PEG) polymer that is coupled to the protein A/G molecule;
- and
- a composition configured to attach a fluorescent label to an antigen protein.
Dependencies are inferred from the claim text.