Evaluating the equilibrium association constant between artinM lectin and myeloid leukemia cells by impedimetric and piezoelectric label free approaches

Label-free methods for evaluating lectin-cell binding have been developed to determine the lectin-carbohydrate interactions in the context of cell-surface oligosaccharides. In the present study, mass loading and electrochemical transducer signals were compared to characterize the interaction between...

Descripción completa

Detalles Bibliográficos
Autores: Carvalho, Fernanda C. [UNESP], Martins, Denise C. [UNESP], Santos, Adriano [UNESP], Roque-Barreira, Maria-Cristina, Bueno, Paulo R. [UNESP]
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2014
País:Brasil
Institución:Universidade Estadual Paulista (UNESP)
Repositorio:Repositório Institucional da UNESP
Idioma:inglés
OAI Identifier:oai:repositorio.unesp.br:11449/131061
Acceso en línea:http://dx.doi.org/10.3390/bios4040358
http://hdl.handle.net/11449/131061
Access Level:acceso abierto
Palabra clave:Artinm
Langmuir isotherm
Electrochemical impedance spectroscopy
Equilibrium association constant (ka)
Lectin
Myeloid leukemia cells
Quartz crystal microbalance
Descripción
Sumario:Label-free methods for evaluating lectin-cell binding have been developed to determine the lectin-carbohydrate interactions in the context of cell-surface oligosaccharides. In the present study, mass loading and electrochemical transducer signals were compared to characterize the interaction between lectin and cellular membranes by measuring the equilibrium association constant, Ka , between ArtinM lectin and the carbohydrate sites of NB4 leukemia cells. By functionalizing sensor interfaces with ArtinM, it was possible to determine Ka over a range of leukemia cell concentrations to construct analytical curves from impedimetric and/or mass-associated frequency shifts with analytical signals following a Langmuir pattern. Using the Langmuir isotherm-binding model, the Ka obtained were (8.9 ± 1.0) × 10(-5) mL/cell and (1.05 ± 0.09) × 10(-6) mL/cell with the electrochemical impedance spectroscopy (EIS) and quartz crystal microbalance (QCM) methods, respectively. The observed differences were attributed to the intrinsic characteristic sensitivity of each method in following Langmuir isotherm premises.