Compositions and methods for making antibody conjugates
The invention relates to antibody conjugates (e.g., a bispecific antibody), drug and nanoparticle compositions and methods and compositions for generating them. This invention further relates to methods of using these compositions for imaging, diagnosing or treating a disease.
1. An adapter comprising an antibody binding domain (AbBD), wherein the AbBD is a variant of a Protein G subdomain comprising an amino acid sequence with at least 80% identity to the sequence set forth in SEQ ID NO: 2 and having a photo-reactive amino acid replacement and/or photo-reactive amino acid modification at A24, K28 or both of the sequence with at least 80% identity to the sequence set forth in SEQ ID NO: 2, and wherein the AbBD specifically binds and crosslinks to an immunoglobulin.
2. The adapter of claim 1 , wherein the photo-reactive amino acid replacement is benzoylphenylalanine (BPA).
3. The adapter of claim 1 , wherein one or more cysteines are engineered into the AbBD and is modified with a photoreactive moiety.
4. The adapter according to claim 1 , wherein said AbBD further comprises a recognition motif for expressed protein ligation (EPL).
5. A conjugate composition comprising the adapter of claim 1 , wherein the adapter is conjugated to a protein, a targeting agent of interest, an oligonucleotide, a DNA, an RNA, a detectable label, an imaging agent, a drug, a toxin, a hapten, a chelate, a polymer, a chemical handle, or a combination thereof.
6. The conjugate composition of claim 5 , wherein the chemical handle is a click chemistry group selected from the group consisting of an azide, an alkyne, a constrained alkyne, dibenzocyclooctyne, a tetrazine, and a transcyclooctyne.
7. A fusion protein comprising the adapter of claim 1 fused in frame with an additional amino acid, a peptide, or a protein.
8. The fusion protein of claim 7 , wherein the additional amino acid, peptide, or protein is selected from the group consisting of SpyTag, SpyCatcher, one half of a split intein, an intein, a cysteine, an N-terminal glycine, and a sortase recognition motif.
9. The fusion protein of claim 7 , wherein the AbBD is fused in frame with a fluorescent protein, a β-galactosidase, a chloramphenicol acetyl transferase, or a luciferase.
10. The fusion protein according to claim 7 , further comprising an affinity tag.
11. The fusion protein according to claim 7 , further comprising a recognition motif for expressed protein ligation (EPL).
12. The fusion protein of claim 7 , wherein said er fusion protein is conjugated to a protein, a targeting agent of interest, an oligonucleotide, a DNA, an RNA, an imaging agent, a drug, a toxin, a hapten, a chelate, a polymer, a chemical handle, or a combination thereof.
13. The fusion protein of claim 12 , wherein the chemical handle is a click chemistry group selected from the group consisting of an azide, an alkyne, a constrained alkyne, dibenzocyclooctyne, a tetrazine, and a transcyclooctyne.
14. The fusion protein of claim 7 , wherein said additional amino acid, peptide, or protein is a targeting ligand, is an antibody, or an antibody fragment.
15. A conjugate composition comprising the adapter of claim 1 operably linked to an immunoglobulin Fc region.
16. The conjugate composition of claim 15 , wherein the immunoglobulin is an IgG molecule.