title: Real-time and label-free analysis of binding thermodynamics of carbohydrate-protein interactions on unfixed cancer cell surfaces using a QCM biosensor.
abstract: A novel approach to the study of binding thermodynamics and kinetics of carbohydrate-protein interactions on unfixed cancer cell surfaces using a quartz crystal microbalance (QCM) biosensor was developed, in which binding events take place at the cell surface, more closely mimicking a biologically relevant environment. In this study, colon adenocarcinoma cells (KM-12) and ovary adenocarcinoma cells (SKOV-3) grew on the optimized polystyrene-coated biosensor chip without fixation. The association and dissociation between the cell surface carbohydrates and a range of lectins, including WGA, Con A, UEA-I, GS-II, PNA and SBA, were monitored in real time and without label for evaluation of cell surface glycosylation. Furthermore, the thermodynamic and kinetic parameters of the interaction between lectins and cell surface glycan were studied, providing detailed information about the interactions, such as the association rate constant, dissociation rate constant, affinity constant, as well as the changes of entropy, enthalpy and Gibbs free energy. This application provides an insight into the cell surface glycosylation and the complex molecular recognition on the intact cell surface, which may have impacts on disease diagnosis and drug discovery.
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
authors.forename: Xueming
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
authors.forename: Siyu
authors.initials: S
authors.lastname: Song
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Shuai
authors.affiliation: Clare College, Cambridge CB2 1TL, United Kingdom.
authors.forename: Yihan
authors.initials: Y
authors.lastname: Pei
authors.affiliation: Attana, SE-11419, Stockholm, Sweden.
authors.forename: Teodor
authors.initials: T
authors.lastname: Aastrup
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Pei
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.|Attana, SE-11419, Stockholm, Sweden.
authors.forename: Zhichao
authors.initials: Z
authors.lastname: Pei
mesh_terms: D015374:Biosensing Techniques; D002241:Carbohydrates / Q000737:chemistry; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000378:metabolism; D009369:Neoplasms / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D058525:Quartz Crystal Microbalance Techniques; D013816:Thermodynamics
chemical_list: D002241:Carbohydrates; D037102:Lectins; D011506:Proteins