Site specific ligation of proteins to synthetic particles

• US 20020187198A1
• Filed: 10/18/2001
• Published: 12/12/2002
• Est. Priority Date: 04/13/1999
• Status: Active Grant

• Current Assignee: Lee, Stephen C.
• Original Assignee: Lee, Stephen C.
• Inventors: Lee, Stephen C.
• Granted Patent: US 6,682,727 B2
• Time in Patent Office: Days
• US Class Current: 424/491
• CPC Class Codes: A61K 47/595 Polyamides, e.g. nylon polyamino acids A61K47/62
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 1/1077 by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids

Abstract

The present invention relates to site-specific conjugation of synthetic particles to proteins.

Claims

1. A substantially homogenous conjugate composed of a synthetic particle attached to the N-terminous of a protein.

2. The conjugate of claim 1 wherein the synthetic particle is a monodisperse synthetic particle.

3. The conjugate of claim 1 wherein the synthetic particle is a monodisperse nanoscale particle.

4. The conjugate of claim 1 wherein the synthetic particle is a dendrimer.

5. The conjugate of claim 1 wherein the synthetic particle is a PAMAM dendrimer.

6. The conjugate of claim 1, 2, 3 or 4 wherein the protein is a four-helical bundle protein.

7. The conjugate of claim 1, 2, 3 or 4 wherein the protein is a cytokine.

8. A pharmaceutical composition of the conjugate of claim 1, 2, 3, 4, 5, 6 or 7.

9. A method for site specifically attaching a synthetic particle to the N-terminus of a protein, said method comprising the steps of:

a) attaching a spacer on the N-terminus of a protein;

b) forming a sulfhydryl on a synthetic particle at the amine; and

c) combining said sulfhydrylized synthetic particle to said spacer on the protein.

10. A method for site specifically attaching a synthetic particle to the N-terminus of a protein, said method comprising the steps of:

a) converting a ser-terminated protein to an aldehyde;

b) converting the amine of a synthetic particle to a oxiamine; and

c) combining said aldehyde ser-terminated protein to said synthetic particle at said oxiamine.

11. The method of claim 9 or 10 wherein the synthetic particle is a monodisperse synthetic particle.

12. The method of claim 9 or 10 wherein the synthetic particle is a PAMAM dendrimer.

13. The method of claim 9 or 10 wherein the synthetic particle is a highly monodisperse nanoscale particle.

14. The method of claim 9 or 10 wherein the synthetic particle is a dendrimer.

15. The method of claim 9, 10, 11, 12, 13 or 14 wherein the protein is a four-helical bundle protein.

16. The method of claim 9, 10, 11, 12, 13 or 14 wherein the protein is a cytokine.