TY - JOUR
T1 - Site-specific chemical modification of recombinant proteins produced in mammalian cells by using the genetically encoded aldehyde tag
AU - Wu, Peng
AU - Shui, Wenqing
AU - Carlson, Brian L.
AU - Hu, Nancy
AU - Rabuka, David
AU - Lee, Julia
AU - Bertozzi, Carolyn R.
PY - 2009/3/3
Y1 - 2009/3/3
N2 - The properties of therapeutic proteins can be enhanced by chemical modification. Methods for site-specific protein conjugation are critical to such efforts. Here, we demonstrate that recombinant proteins expressed in mammalian cells can be site-specifically modified by using a genetically encoded aldehyde tag. We introduced the peptide sequence recognized by the endoplasmic reticulum (ER)-resident formylglycine generating enzyme (FGE), which can be as short as 6 residues, into heterologous proteins expressed in mammalian cells. Cotranslational modification of the proteins by FGE produced products bearing a unique aldehyde group. Proteins bearing this "aldehyde tag" were chemically modified by selective reaction with hydrazide- or aminooxy-functionalized reagents. We applied the technique to site-specific modification of monoclonal antibodies, the fastest growing class of biopharmaceuticals, as well as membrane-associated and cytosolic proteins expressed in mammalian cells.
AB - The properties of therapeutic proteins can be enhanced by chemical modification. Methods for site-specific protein conjugation are critical to such efforts. Here, we demonstrate that recombinant proteins expressed in mammalian cells can be site-specifically modified by using a genetically encoded aldehyde tag. We introduced the peptide sequence recognized by the endoplasmic reticulum (ER)-resident formylglycine generating enzyme (FGE), which can be as short as 6 residues, into heterologous proteins expressed in mammalian cells. Cotranslational modification of the proteins by FGE produced products bearing a unique aldehyde group. Proteins bearing this "aldehyde tag" were chemically modified by selective reaction with hydrazide- or aminooxy-functionalized reagents. We applied the technique to site-specific modification of monoclonal antibodies, the fastest growing class of biopharmaceuticals, as well as membrane-associated and cytosolic proteins expressed in mammalian cells.
KW - Antibody engineering
KW - Bioorthogonal reaction
UR - http://www.scopus.com/inward/record.url?scp=62549121136&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=62549121136&partnerID=8YFLogxK
U2 - 10.1073/pnas.0807820106
DO - 10.1073/pnas.0807820106
M3 - Article
C2 - 19202059
AN - SCOPUS:62549121136
SN - 0027-8424
VL - 106
SP - 3000
EP - 3005
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 9
ER -