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) that specifically binds and crosslinks to an immunoglobulin, wherein the AbBD is a variant of a Protein G subdomain comprising an amino acid sequence with at least 90% identity to the sequence set forth in SEQ ID NO: 2 and having an amino acid replacement and/or an amino acid modification at A24, K28 or both of SEQ ID NO: 2 that is photo-reactive.
2. The adapter of claim 1 , wherein the photo-reactive amino acid replacement is benzoylphenylalanine (BPA).
3. An adapter comprising an antibody binding domain (AbBD) that specifically binds and crosslinks to an immunoglobulin, wherein the AbBD is a variant of a protein G subdomain comprising the amino acid sequence as set forth in SEQ ID NO: 2 and having one or more photo-reactive non-natural amino acids that are incorporated into the AbBD, and wherein the photo-reactive amino acid is benzoylphenylalanine (BPA).
4. The adapter of claim 3 , wherein the BPA replaces A24 or K28 of SEQ ID NO: 2.
5. The adapter of claim 1 , wherein a cysteine is engineered into the AbBD and is modified with a photo-reactive moiety.
6. The adapter according to claim 1 , wherein said AbBD further comprises a recognition motif for expressed protein ligation (EPL).
7. 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.
8. The conjugate composition of claim 7 , 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.
9. A fusion protein comprising the adapter of claim 1 fused in frame with an additional amino acid, a peptide or a protein.
10. The fusion protein of claim 9 , wherein the additional amino acid, peptide, or protein is selected from the group consisting of SpyTag, SpyCatcher, one half of a split intein, an intent, a cysteine, an N-terminal glycine, and a sortase recognition motif.
11. The fusion protein of claim 9 , wherein the AbBD is fused in frame with a fluorescent protein, a β-galactosidase, a chloramphenicol acetyl transferase, or a luciferase.
12. The fusion protein according to claim 9 , further comprising an affinity tag.
13. The fusion protein according to claim 9 , further comprising a recognition motif for expressed protein ligation (EPL).
14. The fusion protein of claim 9 wherein said fusion protein is conjugated to a or 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.
15. The fusion protein of claim 14 , 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.
16. The fusion protein of claim 9 , wherein said additional amino acid, peptide, or protein is a targeting ligand, an antibody, or an antibody fragment.
17. A conjugate composition comprising the adapter of claim 1 operably linked to an immunoglobulin Fc region.
18. The conjugate composition of claim 17 , wherein the immunoglobulin is an IgG molecule.