Macromolecules comprising a thioether cross-link
The present invention provides macromolecules comprising at least one thioether cross-link. A thioether cross-link comprising a single thioether bond between two residues of a macromolecule. The macromolecules of the invention can display enhanced stability, pharmaceutical properties and functional properties. In particular, the invention provides an isolated antibodies comprising at least one thioether cross-link that specifically bind to particular antigens. The present invention also provides a composition comprising a macromolecule substantially free of a denaturing reagent, wherein the macromolecule comprises at least one thioether cross-link. In addition, the present invention provides a method for producing the macromolecules and compositions of the invention.
1. A composition substantially free of a denaturing reagent comprising a monoclonal antibody population and a carrier, wherein each monoclonal antibody in the population comprises the same amino acid sequence, wherein about 0.05%-4% of said population comprises one or multiple thioether cross-link, wherein the thioether cross-link links a heavy chain of the monoclonal antibody and a light chain of the monoclonal antibody, wherein the monoclonal antibody is capable of specifically binding to a respiratory syncytial virus (RSV) antigen provided that the monoclonal antibody is not palivizumab, and wherein said monoclonal antibody population is an IgG 1 .
2. The composition of claim 1 , wherein less than 2%, 1%, 0.75%, 0.5%, 0.25%, or 0.1% of said population comprises at least one thioether cross-link.
3. The composition of claim 1 , wherein the monoclonal antibody population is a population of a human antibody, a chimeric antibody, and/or a humanized antibody.
4. The composition of claim 1 , wherein said about 0.05%-4% of said population comprises multiple thioether cross-links.
5. The composition of claim 1 , wherein the thioether cross-link is in the Fab portion or the Fc portion of the antibody.
6. The composition of claim 1 , wherein the thioether cross-link links a cysteine residue of the heavy chain and a residue of the light chain.
7. The composition of claim 1 , wherein the thioether links a residue of the heavy chain and a cysteine residue of the light chain of the antibody.
8. The composition of claim 1 , wherein the thioether cross-link links a cysteine residue of the heavy chain and a cysteine residue of the light chain.
9. The composition of claim 7 , wherein the thioether cross-link links a cysteine residue in the C H 1 region of the heavy chain and a cysteine residue in the C L region of the light chain.
10. The composition of claim 9 , wherein the cysteine residue in the C H 1 region of the heavy chain is at the amino acid position 223 according to the Kabat numbering system.
11. The composition of claim 9 , wherein the cysteine residue in the C L region of the light chain is at the amino acid position 213 according to the Kabat numbering system.
12. The composition of claim 1 , wherein the carrier is water.
13. The composition of claim 1 , wherein the monoclonal antibody is motavizumab.
14. The composition of claim 1 , wherein the thioether crosslink is lanthionine.
15. The composition of claim 8 , wherein the cysteine residue in the C H 1 region of the heavy chain is at the amino acid position 223 according to the Kabat numbering system, and the cysteine residue in the C L region of the heavy chain is at the amino acid position 213 according to the Kabat numbering system.
16. The composition of claim 15 , wherein the monoclonal antibody is motavizumab.