Protein interactions with HER-family receptors can have different characteristics             depending on the hosting cell line.
Cell lines are common model systems in the development of therapeutic proteins             and in the research on cellular functions and dysfunctions. In this field, the             protein interaction assay is a frequently used tool for assessing the adequacy             of a protein for diagnostic and therapeutic purposes. In this study, we investigated             the extent to which the interaction characteristics depend on the choice of cell             line for HER-family receptors. The interaction characteristics of two therapeutic             antibodies (trastuzumab and cetuximab) and one Affibody molecule (ZHER2:342),             interacting with the intended receptor were characterized with high precision             using an automated real-time interaction method, in different cell lines (HaCaT,             A431, HEP-G2, SKOV3, PC3, DU-145). Clear differences in binding affinity and kinetics,             up to one order of magnitude, were found for the interaction of the same protein             binding to the same receptor on different cells for all three proteins. For HER-family             receptors, it is therefore important to refer to the measured affinity for a protein-receptor             interaction together with the hosting cell line. The ability to accurately measure             affinity and kinetics of a protein-receptor interaction on cell lines of different             origins may increase the understanding of underlying receptor biology, and impact             the selection of candidates in the development of therapeutic or diagnostic agents.
Department of Pharmacology and Toxicology, Faculty of Pharmacy                     in Hradec Kralove, Charles University in Prague, Heyrovského 1203, 50005                     Hradec Králové, Czech Republic.
Pavel
P
Barta
Jennie
J
Malmberg
Ludmila
L
Melicharova
John
J
Strandgård
Anna
A
Orlova
Vladimir
V
Tolmachev
Milan
M
Laznicek
Karl
K
Andersson
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D061067:Antibodies, Monoclonal, Humanized / Q000378:metabolism*; D000915:Antibody Affinity; D000068818:Cetuximab; D066246:ErbB Receptors / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D007457:Iodine Radioisotopes; D007700:Kinetics; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D011869:Radioligand Assay; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D000068878:Trastuzumab
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000068818:Cetuximab; D066246:ErbB Receptors; D007457:Iodine Radioisotopes; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; D000068878:Trastuzumab; C533351:affibody (ZHER2-342)2; C512478:EGFR protein, human; C508053:ERBB2 protein, human