TY - JOUR
T1 - Small-Molecule Probes for Affinity-Guided Introduction of Biocompatible Handles on Metal-Binding Proteins
AU - Mortensen, Michael Rosholm
AU - Skovsgaard, Mikkel B.
AU - Okholm, Anders Hauge
AU - Scavenius, Carsten
AU - Dupont, Daniel Miotto
AU - Rosen, Christian Bech
AU - Enghild, Jan J.
AU - Kjems, Jørgen
AU - Gothelf, Kurt Vesterager
PY - 2018/9/19
Y1 - 2018/9/19
N2 - Protein conjugates of high heterogeneity may contain species with significantly different biological properties, and as a consequence, the focus on methods for production of conjugates of higher quality has increased. Here, we demonstrate an efficient and generic approach for the modification of metal-binding proteins with biocompatible chemical handles without the need for genetic modifications. Affinity-guided small-molecule probes are developed for direct conjugation to off-the-shelf proteins and for installing different chemical handles on the protein surface. While purification of protein conjugates obtained by small molecule conjugation is troublesome, the affinity motifs of the probes presented here allow for purification of the conjugates. The versatility of the probes is demonstrated by conjugation to several His-tagged and natural metal-binding proteins, including the efficient and area-selective conjugation to three therapeutically relevant antibodies.
AB - Protein conjugates of high heterogeneity may contain species with significantly different biological properties, and as a consequence, the focus on methods for production of conjugates of higher quality has increased. Here, we demonstrate an efficient and generic approach for the modification of metal-binding proteins with biocompatible chemical handles without the need for genetic modifications. Affinity-guided small-molecule probes are developed for direct conjugation to off-the-shelf proteins and for installing different chemical handles on the protein surface. While purification of protein conjugates obtained by small molecule conjugation is troublesome, the affinity motifs of the probes presented here allow for purification of the conjugates. The versatility of the probes is demonstrated by conjugation to several His-tagged and natural metal-binding proteins, including the efficient and area-selective conjugation to three therapeutically relevant antibodies.
U2 - 10.1021/acs.bioconjchem.8b00424
DO - 10.1021/acs.bioconjchem.8b00424
M3 - Journal article
C2 - 30091905
SN - 1043-1802
VL - 29
SP - 3016
EP - 3025
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
IS - 9
ER -