Methods for isolating proteins expressed by dendritic cells
OwnerMark A. Velleca, Ira Mellmaninventors
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Abstract
Disclosed are methods for generating dendritic cells and isolating dendritic cell associated proteins. Also disclosed are compositions and methods for generating an antibody against a dendritic cell associated protein as well as methods for modulating the expression or activity of dendritic cell associated proteins. The compositions and methods of the invention are useful in the diagnosis and treatment of diseases associated with altered dendritic cell activity.
Claims31
A method of isolating a dendritic cell associated protein comprising:
- (a) treating a dendritic cell with a first agent which alters the expression or activity of a protein associated with the dendritic cell;
- (b) isolating a cell extract from the dendritic cell;
- (c) associating the extract with at least one second agent;
- (d) detecting the binding of the second agent to the dendritic cell associated protein in the extract;
- and
- (e) isolating the dendritic cell associated protein bound to the second agent from the extract.
A method of detecting the presence of a disease associated with altered dendritic cell activity in a patient comprising:
- (a) isolating a sample of dendritic cells from the patient;
- (b) associating the sample with an agent which binds to a dendritic cell associated protein;
- (c) measuring the amount of binding of the agent to the dendritic cell associated protein;
- wherein an alteration in the amount of binding is indicative of a disease associated with altered dendritic cell activity.
A method of producing an antibody against a dendritic cell associated protein comprising the steps of:
- (a) isolating the dendritic cell associated protein;
- (b) generating a phage display antibody library against the protein;
- (c) associating the phage library with the dendritic cell associated protein;
- and
- (d) isolating an antibody from the phage display antibody library that binds to the protein.
A method of screening for an agent that modulates the expression or activity of a dendritic cell associated protein comprising:
- (a) contacting a dendritic cell with a first agent which alters the expression or activity of a protein associated with the dendritic cell;
- (b) contacting the dendritic cell with a second agent;
- and
- (c) measuring the level of expression of the dendritic cell associated protein or mRNA encoding the dendritic cell associated protein;
- wherein an alteration in the level of expression indicates that the second agent modulates the expression or activity of the dendritic cell associated protein.
Dependencies are inferred from the claim text.