Site specific ligation of proteins to synthetic particles

• US 6,682,727 B2
• Filed: 10/18/2001
• Issued: 01/27/2004
• Est. Priority Date: 04/13/1999
• Status: Expired due to Fees

• Current Assignee: GD Searle LLC (Pfizer Inc.)
• Original Assignee: GD Searle LLC (Pfizer Inc.)
• Inventors: Parthasarathy, Ranjani V., Lee, Stephen C.
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US09/982,765
• Publication Number:
  US 20020187198A1
• Time in Patent Office: 831 Days
• Field of Search: 424/78.17, 530/300, 530/350
• US Class Current: 424/78.17
• 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 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.

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

3. The method of claim 1 wherein the synthetic particle is a PAMAM dendrimer.

4. The method of claim 1 wherein the synthetic particle is a highly monodisperse nanoscale particle.

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

6. The method of claim 1, 2, 3, 4 or 5 wherein the protein is a four-helical bundle protein.

7. The method of claim 1, 2, 3, 4 or 5 wherein the protein is a cytokine.