Methods of identifying epitopes
AssigneeOblique Therapeutics AB
InventorsOwe ORWAR, Carolina TRKULJA, Max DAVIDSON
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Abstract
The present invention relates to methods of identifying an epitope on a protein that can be bound by an antibody. Methods of the invention typically involve a step of limited or restricted proteolysis of a protein and the identification of sites on the protein that are cut by the protease(s) used. The invention also relates to antibodies which bind to epitopes that have been identified by methods of the invention.
Claims28
A method of identifying an epitope on a protein that can be bound by an antibody, said method comprising:
- performing in silico protease digestion of said protein with one or more proteases to identify sites on the protein that are predicted to be cut by said one or more proteases;
- (ii) performing in vitro limited or restricted proteolysis of said protein with one or more proteases;
- (iii) identifying peptides released from said protein by the in vitro protease digestion of step (ii), thereby identifying cut sites;
- (iv) comparing the in silico predicted cut sites identified in step (i) with cut sites identified in step (iii);
- (v) probing one or more epitopes in a region of the protein containing or flanking a cut site that is an in silico predicted protease cut site identified in step (i) but that is not a cut site identified in step (iii) with one or more antibodies;
- and
- (vi) identifying whether or not said one or more antibodies bind to said one or more epitopes thereby identifying an epitope on a protein that can be bound by an antibody.
A method of identifying an epitope on a protein that can be bound by an antibody, said method comprising:
- identifying sites at which one or more protease has cut said protein after said protein has been exposed to limited or restricted proteolysis by said one or more protease;
- and
- (ii) probing a plurality of epitopes on said protein that are between the cut sites, that overlap with a cut site, or that are in a region that flanks a cut site with antibodies directed to said epitopes, thereby identifying one or more epitopes that can be bound by an antibody.
Dependencies are inferred from the claim text.