title: Study of the interaction of trastuzumab and SKOV3 epithelial cancer cells using a quartz crystal microbalance sensor.
abstract: Analytical methods founded upon whole cell-based assays are of importance in early stage drug development and in fundamental studies of biomolecular recognition. Here we have studied the binding of the monoclonal antibody trastuzumab to human epidermal growth factor receptor 2 (HER2) on human ovary adenocarcinoma epithelial cancer cells (SKOV3) using quartz crystal microbalance (QCM) technology. An optimized procedure for immobilizing the cells on the chip surface was established with respect to fixation procedure and seeding density. Trastuzumab binding to the cell decorated sensor surface was studied, revealing a mean dissociation constant, KD, value of 7 ± 1 nM (standard error of the mean). This study provides a new perspective on the affinity of the antibody-receptor complex presented a more natural context compared to purified receptors. These results demonstrate the potential for using whole cell-based QCM assay in drug development, the screening of HER2 selective antibody-based drug candidates, and for the study of biomolecular recognition. This real time, label free approach for studying interactions with target receptors present in their natural environment afforded sensitive and detailed kinetic information about the binding of the analyte to the target.
authors.affiliation: Bioorganic & Biophysical Chemistry Laboratory, Linnæus University Centre for Biomaterials Chemistry, Department of Chemistry & Biomedical Sciences, Linnæus University, SE-39182 Kalmar, Sweden. louise.elmlund@lnu.se.
authors.forename: Louise
authors.initials: L
authors.lastname: Elmlund
authors.affiliation: Attana AB, Björnnäsvägen 21, SE-11419 Stockholm, Sweden. camilla.kack@attana.com.
authors.forename: Camilla
authors.initials: C
authors.lastname: Käck
authors.affiliation: Attana AB, Björnnäsvägen 21, SE-11419 Stockholm, Sweden. teodor.aastrup@attana.com.
authors.forename: Teodor
authors.initials: T
authors.lastname: Aastrup
authors.affiliation: Bioorganic & Biophysical Chemistry Laboratory, Linnæus University Centre for Biomaterials Chemistry, Department of Chemistry & Biomedical Sciences, Linnæus University, SE-39182 Kalmar, Sweden. ian.nicholls@lnu.se.|Department of Chemistry-BMC, Uppsala University, Box 576, SE-75123 Uppsala, Sweden. ian.nicholls@lnu.se.
authors.forename: Ian A
authors.initials: IA
authors.lastname: Nicholls
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage* / Q000737:chemistry; D015374:Biosensing Techniques; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000187:drug effects / Q000473:pathology; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000473:pathology; D011791:Quartz / Q000737:chemistry; D058525:Quartz Crystal Microbalance Techniques; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D011791:Quartz; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human