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Science 7 February 2003:
Vol. 299. no. 5608, pp. 884 - 887
DOI: 10.1126/science.1079085
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Reports

Design and Chemical Synthesis of a Homogeneous Polymer-Modified Erythropoiesis Protein

Gerd G. Kochendoerfer,1* Shiah-Yun Chen,1 Feng Mao,1 Sonya Cressman,1dagger Stacey Traviglia,1ddagger Haiyan Shao,1 Christie L. Hunter,1§ Donald W. Low,1 E. Neil Cagle,1 Maia Carnevali,1 Vincent Gueriguian,1 Peter J. Keogh,1 Heather Porter,1 Stephen M. Stratton,1 M. Con Wiedeke,1 Jill Wilken,1 Jie Tang,1parallel Jay J. Levy,1 Les P. Miranda,1 Milan M. Crnogorac,1 Suresh Kalbag,1 Paolo Botti,1# Janice Schindler-Horvat,1 Laura Savatski,2 John W. Adamson,2 Ada Kung,1 Stephen B. H. Kent,1** James A. Bradburne1

We report the design and total chemical synthesis of "synthetic erythropoiesis protein" (SEP), a 51-kilodalton protein-polymer construct consisting of a 166-amino-acid polypeptide chain and two covalently attached, branched, and monodisperse polymer moieties that are negatively charged. The ability to control the chemistry allowed us to synthesize a macromolecule of precisely defined covalent structure. SEP was homogeneous as shown by high-resolution analytical techniques, with a mass of 50,825 ±10 daltons by electrospray mass spectrometry, and with a pI of 5.0. In cell and animal assays for erythropoiesis, SEP displayed potent biological activity and had significantly prolonged duration of action in vivo. These chemical methods are a powerful tool in the rational design of protein constructs with potential therapeutic applications.

1 Gryphon Therapeutics, 250 East Grand Avenue, Suite 90, South San Francisco, CA 94080, USA.
2 The Blood Research Institute, Blood Center of Southeastern Wisconsin, 8727 Watertown Plank Road, Milwaukee, WI 53226, USA.
*   To whom correspondence should be addressed. E-mail: Gkochendoerfer{at}gryphonRX.com

dagger    Present address: Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of British Columbia, 2146 Health Sciences Mall, Vancouver, BC V6T 1Z3, Canada.

ddagger    Present address: Diosynth RTP, Inc., 3000 Weston Parkway, Cary, NC 27513, USA.

§   Present address: Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404, USA.

parallel    Present address: Celera Genomics, 180 Kimball Way, South San Francisco, CA 94080, USA.

   Present address: Midwest Biotech, 12690 Ford Drive, Fishers, IN 46038-1151, USA.

#   Present address: GeneProt Inc., 2 Pré-de-la-Fontaine, 1217 Meyrin/Geneva, Switzerland.

**   Present address: Departments of Biochemistry & Molecular Biology, and The University of Chicago, Chicago, IL 60637, USA.

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