Evaluating the Equilibrium Association Constant between ArtinM Lectin and Myeloid Leukemia Cells by Impedimetric and Piezoelectric Label Free Approaches

Abstract

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.

The lectin ArtinM activates RBL-2H3 mast cells without inducing degranulation

“…The α subunit of FcεRI, which is exposed on the cell surface, is highly glycosylated [58]. ArtinM is a homotetramer with 4 carbohydrate recognition domains that is capable of binding to and cross-linking cell surface carbohydrates [59]. It is possible that in low concentrations, the degree of cross-linking may be sufficient to activate mast cells, but insufficient to stimulate degranulation.…”

Recent Progress in Electrochemical Biosensors for Glycoproteins

“…It is possible to use SAM-coated Au electrodes modified with an oligosaccharide to detect lectin and influenza hemagglutinin [71,72]. Bueno and coworkers have prepared lectin-immobilized sensors for studying lectin-glycoprotein interactions based on the impedimetric and capacitive responses of the sensors [73,74]. For this purpose, ArtinM lectin was covalently immobilized on a SAM-coated electrode and the binding behavior to HRP and glycoprotein-bearing leukemia cells was studied.…”

Glycoprotein assay based on the optimized immittance signal of a redox tagged and lectin-based receptive interface

“…Indeed several cancer protein biomarkers (proteins related to trauma, infections, and disease onset or progression (Mayeux, 2004;Strimbu and Tavel, 2010)) are glycoproteins presenting with aberrant changes in carbohydrate content such as the prostatic specific antigen (PSA, for prostatic cancer) and CA125 (for ovarian cancer) (Adamczyk et al, 2012;Silva, 2015). Accordingly, the methods for both carbohydrate/glycoprotein detection and qualification/quantification of carbohydrate/glycoprotein-protein/ cell interaction are greatly studied in literature (Pierce et al, 1999;Laurent et al, 2008;Yakovleva et al, 2010;Safina et al, 2011;Bertok et al, 2013aBertok et al, , 2013bCarvalho et al, 2014;Santos et al, 2014a;Surinova et al, 2015).…”

“…It allows detection of changes in the optical properties as a consequence of protein adsorption that can be applied in affinity studies (Homola et al, 1999;Laurent et al, 2008;Safina et al, 2011). Nonetheless, the development of glycoarrays based either QCM or SPR is time-consuming, presenting with higher cost and lower sensitivity compared to competing electroanalytical techniques such as impedance electrochemical assays (Carvalho et al, 2014;Santos et al, 2014a).…”