Design and chemical synthesis of a homogeneous polymer-modified erythropoiesis protein.
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.
Kochendoerfer
Gerd G
GG
Gryphon Therapeutics, 250 East Grand Avenue, Suite 90, South San Francisco, CA 94080, USA. Gkochendoerfer@gryphonRX.com
Chen
Shiah-Yun
SY
Mao
Feng
F
Cressman
Sonya
S
Traviglia
Stacey
S
Shao
Haiyan
H
Hunter
Christie L
CL
Low
Donald W
DW
Cagle
E Neil
EN
Carnevali
Maia
M
Gueriguian
Vincent
V
Keogh
Peter J
PJ
Porter
Heather
H
Stratton
Stephen M
SM
Wiedeke
M Con
MC
Wilken
Jill
J
Tang
Jie
J
Levy
Jay J
JJ
Miranda
Les P
LP
Crnogorac
Milan M
MM
Kalbag
Suresh
S
Botti
Paolo
P
Schindler-Horvat
Janice
J
Savatski
Laura
L
Adamson
John W
JW
Kung
Ada
A
Kent
Stephen B H
SB
Bradburne
James A
JA
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002942:Circular Dichroism; D015195:Drug Design; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D004920:Erythropoiesis; D004921:Erythropoietin / Q000737:chemistry / Q000494:pharmacology; D006400:Hematocrit; D006801:Humans; D007526:Isoelectric Point; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D011108:Polymers / Q000138:chemical synthesis* / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D017510:Protein Folding; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D051381:Rats; D017467:Receptors, Erythropoietin / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins; D021241:Spectrometry, Mass, Electrospray Ionization
D011108:Polymers; D011506:Proteins; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; C472014:synthetic erythropoiesis protein; D004921:Erythropoietin