Antibodies comprising site-specific non-natural amino acid residues, methods of their preparation and methods of their use
Inventors
Thanos, Christopher D. • McEvoy, Leslie • Yin, Gang • Penta, Kalyani • Baliga, Ramesh • BAJAD, Sunil • Pollitt, Sonia • Murray, Chris • Steiner, Alex • Gill, Avinash
Assignees
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Publication Number
US-11958909-B2
Publication Date
2024-04-16
Expiration Date
Abstract
Provided herein are antibodies comprising non-natural amino acid residues at site-specific positions, compositions comprising the antibodies, methods of their production and methods of their use. The antibodies are useful for methods of treatment and prevention, methods of detection and methods of diagnosis.
Core Innovation
The invention provides a method of detecting an antigen by contacting a sample comprising the antigen with an antibody conjugate or a pharmaceutical composition comprising the antibody conjugate. The antibody conjugate comprises an isolated antibody of the IgG1 class with a heavy chain and a light chain that binds to the antigen, and the antigen-antibody complex is detected.
The antibody conjugate includes non-natural amino acid residues at specific sites in the heavy chain or light chain according to the Kabat numbering scheme and/or EU index of Kabat. For the heavy chain, the non-natural amino acid residue is positioned at heavy chain residues 19, 25, 40, 52, and 70 according to the Kabat numbering scheme, and at heavy chain residues 119, 121, 180, 190, 222, 241, and 404 according to the EU index of Kabat. For the light chain, the non-natural amino acid residue is positioned at light chain residues 7, 22, and 152.
Each non-natural amino acid residue is independently selected from ortho-substituted tyrosine, meta-substituted tyrosine, para-substituted phenylalanine, ortho-substituted phenylalanine, and meta-substituted phenylalanine. At least one of the non-natural amino acids is linked to a payload capable of being detected, and the antibody conjugate may include an aglycosylated variant.
The document further describes hydroxylamine/oxime-bearing sidechains and conjugate formation involving oxime/hydroxylamine reactions between hydroxylamine/oxime-bearing sidechains and carbonyl-containing or dicarbonyl-containing reagents, to form conjugates or oximes. It additionally includes aqueous stability of Formula (I) under mildly acidic conditions and embodiments in which tRNA recognizes selector codons, including amber codon, ochre codon, opal codon, rare codons, unnatural codons, five-base codons, and four-base codons.
Claims Coverage
The claim coverage centers on antigen detection using an IgG1 antibody conjugate in which substituted tyrosine/phenylalanine non-natural amino acid residues are placed at specified heavy-chain and/or light-chain Kabat positions, at least one of which is linked to a detectable payload, followed by detecting the antigen-antibody complex. The inventive features include site-specific residue selection, defined non-natural amino acid identities, payload linkage, and complex detection.
IgG1 antibody conjugate for antigen detection
A method of detecting an antigen by contacting a sample comprising the antigen with an antibody conjugate or a pharmaceutical composition comprising the antibody conjugate, wherein the antibody conjugate comprises an isolated antibody of the IgG1 class comprising a heavy chain and a light chain that binds to the antigen, and detects a complex formed between the antibody and the antigen.
Heavy-chain site-specific non-natural amino acid residues by Kabat and EU index
The heavy chain comprises at least one non-natural amino acid residue at a specific site selected from heavy chain residues 19, 25, 40, 52, and 70 according to the Kabat numbering scheme and heavy chain residues 119, 121, 180, 190, 222, 241, and 404 according to the EU index of Kabat.
Light-chain site-specific non-natural amino acid residues by Kabat numbering
The light chain comprises a non-natural amino acid residue at a specific site selected from light chain residues 7, 22, and 152 according to the Kabat numbering scheme.
Detectable payload linked to non-natural amino acids
Each non-natural amino acid residue is independently selected from ortho-substituted tyrosine, meta-substituted tyrosine, para-substituted phenylalanine, ortho-substituted phenylalanine, and meta-substituted phenylalanine, wherein at least one of the non-natural amino acids is linked to a payload capable of being detected.
Aglycosylated variant
The method includes a combination of such sites, or an aglycosylated variant thereof.
The claims are directed to antigen detection with an IgG1 antibody conjugate bearing substituted tyrosine/phenylalanine non-natural amino acids at defined Kabat and EU index sites, with at least one residue linked to a detectable payload, and include aglycosylated variants.
Stated Advantages
Aqueous stability of Formula (I) under mildly acidic conditions (pH 2 to 8).
Documented Applications
Detection of an antigen by contacting a sample comprising the antigen with an antibody conjugate or a pharmaceutical composition comprising the antibody conjugate.
Embodiments in which tRNA recognizes selector codons, including amber codon, ochre codon, opal codon, rare codons, unnatural codons, five-base codons, and four-base codons.
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