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US 2026/0062469 A1Published application
Compositions and methods for linear and conformational site-specific antibodies and methods of making the same
OwnerMichael P. Weinerinventor
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Abstract
The present disclosure provides a method of making a site-specific antibody to a target of interest.
Claims30
An antibody prepared according to the process comprising:
- a. immunizing an animal at least once with a modified peptide having an amino acid sequence of about 10 to about 20 amino acids that is identical to an amino acid sequence of a protein of interest except wherein one internal native amino acid has been substituted with a non-native amino acid (nnAA), wherein the nnAA is a phosphorylated amino acid, an acetylated amino acid, an isocyanated amino acid, a sulfated amino acid, or a nitrated amino acid;
- b. boosting the animal at least once with a first unmodified peptide comprising a core amino acid sequence that is identical to the modified peptide of step (a), except wherein the nnAA has been substituted with the native amino acid, a first N-terminal amino acid sequence that is not native to the protein of interest, and a first C-terminal amino acid sequence that is not native to the protein of interest;
- c. cloning B-cells obtained from the animal;
- and
- d. identifying a B-cell clone of step (c) that produce an antibody that:
- i. binds to the first unmodified peptide and a second unmodified peptide comprising
- the core amino acid sequence of the first unmodified peptide,
- a second N-terminal amino acid sequence that is not native to the protein of interest, and
- a second C-terminal amino acid sequence that is not native to the protein of interest,
- wherein the first N-terminal amino acid sequence, the first C-terminal amino acid sequence, the second N-terminal amino acid sequence, and the second C-terminal amino acid sequences are different amino acid sequences.
7 more elements
A method of producing a site directed binding agent to a protein of interest, the method comprising:
- a. providing a modified peptide having an amino acid sequence of about 10 to about 20 amino acids that is identical to an amino acid sequence of a protein of interest except wherein one internal amino acid has been substituted with a non-native amino acid (nnAA), wherein the nnAA is a phosphorylated amino acid, an acetylated amino acid, an isocyanated amino acid, a sulfated amino acid, or a nitrated amino acid;
- b. screening the modified peptide against a library;
- c. isolating one or more binding agents that bind to the modified peptide;
- d. generating a library of clonotypes of the one or more binding agents isolated in step (c);
- e. screening the library of clonotypes against:
- i. the modified peptide of step (a);
- and
- ii. a first unmodified peptide comprising a core amino acid sequence that is identical to the modified peptide of step (a), except wherein the nnAA has been substituted with the native amino acid, a first N-terminal amino acid sequence that is not native to the protein of interest, and a first C-terminal amino acid sequence that is not native to the protein of interest;
- and
- isolating a binding agent that bind to both the first unmodified peptide and the modified peptide.
7 more elements
A kit comprising:
- a. a first compartment comprising a modified peptide having an amino acid sequence of about 10 to about 20 amino acids that is identical to an amino acid sequence of a protein of interest except wherein one internal amino acid has been substituted with a non-native amino acid (nnAA), wherein the nnAA is a phosphorylated amino acid, an acetylated amino acid, an isocyanated amino acid, a sulfated amino acid, or a nitrated amino acid;
- b. a second compartment comprising a first unmodified peptide having a core amino acid sequence that is identical to the modified peptide of step (a), except wherein the nnAA has been substituted with the native amino acid (nAA) and comprising a first N-terminal amino acid sequence and a first C-terminal amino acid sequence that is not native to the protein of interest;
- c. a third compartment comprising a second unmodified peptide having a core amino acid sequence that is identical to the first unmodified peptide and comprising a second N-terminal amino acid sequence that is not native to the protein of interest and a second C-terminal amino acid sequence that is not native to the protein of interest;
- and
- d. instructions for use.
2 more elements
Dependencies are inferred from the claim text.