Antibody conjugates and methods of making and using the same
Antibodies that include a sulfatase motif-containing tag in a constant region of an immunoglobulin (Ig) heavy chain polypeptide are disclosed. The sulfatase motif can be converted by a formylglycine-generating enzyme (FGE) to produce a formylglycine (fGly)-modified Ig heavy chain polypeptide. An fGly-modified Ig heavy chain polypeptide of the antibody can be covalently and site-specifically bound to a moiety of interest to provide an antibody conjugate. The disclosure also encompasses methods of production of such tagged Ig heavy chain polypeptides, fGly-modified Ig heavy chain polypeptides, and antibody conjugates, as well as methods of use of same.
1. An antibody comprising an immunoglobulin (Ig) heavy chain polypeptide and an Ig light chain polypeptide, wherein:
the Ig heavy chain polypeptide has 97% or more sequence identity to the amino acid sequence set forth in SEQ ID NO:64, and
the Ig heavy chain polypeptide comprises an insertion of a sulfatase motif (LCTPSR (SEQ ID NO: 561)) before the asparagine residue at the 91 st position of the amino acid sequence set forth in SEQ ID NO: 1 thereby producing in the Ig heavy chain polypeptide the amino acid sequence of KPSX 1 Z 1 X 2 Z 2 X 3 Z 3 NTK (SEQ ID NO: 246), wherein X 1 is L, Z 1 is C, X 2 is T, Z 2 is P, X 3 is S, and Z 3 is R.
2. The antibody of claim 1 , wherein Ig heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 64.
3. The antibody of claim 1 , wherein, in the Ig heavy chain polypeptide, the cysteine residue in the sulfatase motif (LCTPSR (SEQ ID NO: 561)) is oxidized to fGly.
4. The antibody of claim 3 , wherein, in the Ig heavy chain polypeptide, fGly is covalently bound to a payload.
5. The antibody of claim 4 , wherein the Ig heavy chain polypeptide is covalently bound to the payload via a hydrazone, oxime, semicarbazone, alkyl, alkenyl, acyloxy, hydrazinyl-indolyl, hydrazinyl-imidazoyl, hydrazinyl-pyrrolyl, hydrazinyl-furanyl or a pyrazalinone linkage.
6. The antibody of claim 4 , wherein the Ig heavy chain polypeptide is covalently bound to the payload via a linking group.
7. The antibody of claim 6 , wherein the linking group comprises a 4-aminopiperidine derivative (4AP).
8. The antibody of claim 4 , wherein the payload is selected from a drug, a detectable label, a water-soluble polymer, and a synthetic peptide.
9. The antibody of claim 4 , wherein the payload is a small molecule drug.
10. The antibody of claim 9 , wherein the small molecule drug is a cancer chemotherapeutic agent selected from the group consisting of an alkylating agent, a nitrosourea, an antimetabolite, an antitumor antibiotic, a vinca alkaloid, and a steroid hormone.
11. The antibody of claim 8 , wherein the water-soluble polymer is poly(ethylene glycol).
12. The antibody of claim 8 , wherein the detectable label is an imaging agent.
13. The antibody of claim 4 , wherein the payload is a viral fusion inhibitor.
14. An antibody comprising an immunoglobulin (Ig) heavy chain polypeptide and an Ig light chain polypeptide, wherein:
the Ig heavy chain polypeptide has 97% or more sequence identity to the amino acid sequence set forth in SEQ ID NO: 85, and
the Ig heavy chain polypeptide comprises an insertion of a sulfatase motif (LCTPSR (SEQ ID NO: 561)) before the glutamic acid residue at the 116 th position of the amino acid sequence set forth in SEQ ID NO: 1 thereby producing in the Ig heavy chain polypeptide the amino acid sequence of CPAPX 1 Z 1 X 2 Z 2 X 3 Z 3 E[L/F]L (SEQ ID NO: 268), wherein X 1 is L, Z 1 is C, X 2 is T, Z 2 is P, X 3 is S, and Z 3 is R.
15. The antibody of claim 14 , wherein Ig heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 85.
16. The antibody of claim 14 , wherein, in the Ig heavy chain polypeptide, the cysteine residue in the sulfatase motif (LCTPSR (SEQ ID NO: 561)) is oxidized to fGly.
17. The antibody of claim 16 , wherein, in the Ig heavy chain polypeptide, fGly is covalently bound to a payload.
18. The antibody of claim 17 , wherein the Ig heavy chain polypeptide is covalently bound to the payload via a hydrazone, oxime, semicarbazone, alkyl, alkenyl, acyloxy, hydrazinyl-indolyl, hydrazinyl-imidazoyl, hydrazinyl-pyrrolyl, hydrazinyl-furanyl or a pyrazalinone linkage.
19. The antibody of claim 17 , wherein the Ig heavy chain polypeptide is covalently bound to the payload via a linking group.
20. The antibody of claim 19 , wherein the linking group comprises a 4-aminopiperidine derivative (4AP).
21. The antibody of claim 17 , wherein the payload is selected from a drug, a detectable label, a water-soluble polymer, and a synthetic peptide.
22. The antibody of claim 17 , wherein the payload is a small molecule drug.
23. The antibody of claim 22 , wherein the small molecule drug is a cancer chemotherapeutic agent selected from the group consisting of an alkylating agent, a nitrosourea, an antimetabolite, an antitumor antibiotic, a vinca alkaloid, and a steroid hormone.
24. The antibody of claim 21 , wherein the water-soluble polymer is poly(ethylene glycol).
25. The antibody of claim 21 , wherein the detectable label is an imaging agent.
26. The antibody of claim 17 , wherein the payload is a viral fusion inhibitor.