Alexander S Hoffman

US Patent:

7037894, May 2, 2006

Filed:

Apr 18, 2001

Appl. No.:

09/837235

Inventors:

Christopher P. Marshall - Brooklyn NY, US
Alexander Hoffman - Los Angeles CA, US
Joseph P. Errico - Far Hills NJ, US
Paul B. Marshall - Munich, DE

Assignee:

Avatar Medical, LLC - Newark NJ

International Classification:

A61K 38/00
A61K 38/43
C07K 1/00
C07K 16/00
C12N 9/00

US Classification:

514 12, 424 941, 4241301, 424 943, 530350, 5303871, 53038821, 53038822, 53038824, 530399, 435183, 435198, 514 2

Abstract:

Isolated polypeptides or polypeptide chains are modified by di-tyrosine cross-linking such that the retain at least one functional activity. In one embodiment, the isolated polypeptide or polypeptide chains comprise at least one di-tyrosine cross-link, wherein at least one tyrosine of the di-tyrosine cross-link originates from a point mutation to tyrosine, and wherein the di-tyrosine cross-linked protein retains at least one function displayed by the protein in the absence of di-tyrosine cross-linking. In another embodiment, the di-tyrosine cross-linked polypeptide or polypeptide chain has enhanced stability compared to the same polypeptide or polypeptide chain in the absence of di-tyrosine cross-linking. A method for stabilization of a polypeptide or polypeptide complex, by the introduction of intra-polypeptide and/or inter-polypeptide di-tyrosine bonds, which simultaneously maintains the structure and function of the polypeptide or polypeptide complex is also described.

US Patent:

7445912, Nov 4, 2008

Filed:

Jan 29, 2007

Appl. No.:

11/699588

Inventors:

Christopher P. Marshall - Brooklyn NY, US
Alexander Hoffman - La Jolla CA, US
Joseph P. Errico - Palo Alto CA, US
Paul B. Marshall - Munich, DE

Assignee:

Avatar Medical LLC - New York NY

International Classification:

C12P 21/06
C07K 1/00
C07K 14/00

US Classification:

435 681, 530350, 530300

Abstract:

Isolated polypeptides or polypeptide chains are modified by di-tyrosine cross-linking such that they retain at least one functional activity. In one embodiment, the isolated polypeptide or polypeptide chains comprise at least one di-tyrosine cross-link, wherein at least one tyrosine of the di-tyrosine cross-link originates from a point mutation to tyrosine, and wherein the di-tyrosine cross-linked protein retains at least one function displayed by the protein in the absence of di-tyrosine cross-linking. In another embodiment, the di-tyrosine cross-linked polypeptide or polypeptide chain has enhanced stability compared to the same polypeptide or polypeptide chain in the absence of di-tyrosine cross-linking. A method for stabilization of a polypeptide or polypeptide complex, by the introduction of intra-polypeptide and/or inter-polypeptide di-tyrosine bonds, which simultaneously maintains the structure and function of the polypeptide or polypeptide complex is also described.

US Patent:

20050123530, Jun 9, 2005

Filed:

Jan 26, 2005

Appl. No.:

11/043731

Inventors:

Christopher Marshall - Brooklyn NY, US
Alexander Hoffman - Los Angeles CA, US
Joseph Errico - Palo Alto CA, US
Paul Marshall - Munich, DE

Assignee:

Avatar Medical, LLC - Newark NJ

International Classification:

A61K038/46
A61K039/395

US Classification:

424094600, 435198000, 530391100, 530350000, 530399000, 530400000, 514012000, 424178100

Abstract:

The invention described herein comprises methods for stabilizing polypeptides and polypeptide complexes, and the polypeptides and polypeptide complexes stabilized using the methods. To achieve stabilization, a cross-link reaction is controlled such that polypeptides and polypeptide complexes maintain their original functionality. In one embodiment, the invention provides a method for the identification of amino acid residues which, when cross-linked, are least disruptive to the structure and function of the polypeptide or polypeptide complex. In another embodiment, the invention provides a method for mutagenesis of identified residues to further control the cross-link reaction. Polypeptides and polypeptide complexes so stabilized can be utilized under a wide variety of physiological and non-physiological conditions. Further, the cross-link methodology disclosed herein may preclude the need for addition of exogenous structures to engineered proteins and complexes, such as peptide linkers that could be immunogenic and/or significantly decrease efficacy. In another embodiment, the invention provides a method for statistical analysis of databases of structural and/or sequence information available for polypeptides and polypeptide complexes to be stabilized. The statistical analysis identifies suitable residue pairs which are least likely to be disruptive of structure and function when cross-linked. Further, in a polypeptide chain or chains to be cross-linked, potentially undesirable reactive side-chains may be masked and protected, or altered using site-directed mutagenesis, e.g., to introduce a maximally conservative point mutation that will not support the cross-link reaction. The cross-link reaction conditions may also be adjusted to prevent undesired cross-links or other undesired side-effects. At residues identified as desirable positions for cross-linking, reactive side-chains may be introduced by site-directed mutagenesis, and the cross-link reaction is carried out using the conditions identified above.