Sorption of his-tagged Protein G and Protein G onto chitosan/divalent metal ion sorbent used for detection of microcystin-LR

• Published in: Environmental science and pollution research international 2017, Vol.24 (1), p.15-24
• Main Authors: Demey, Hary, Tria, Scherrine A., Soleri, Romain, Guiseppi-Elie, Anthony, Bazin, Ingrid
• Format: Article
• Language: English
• Subjects: Adsorption; Antibodies, Immobilized; Aquatic Pollution; Atmospheric Protection/Air Quality Control/Air Pollution; Calibration; Chemiluminescence; Chitosan - chemistry; Cyanobacteria; Detection limits; Drinking water; Earth and Environmental Science; Ecotoxicology; Engineering Sciences; Environment; Environmental Chemistry; Environmental Health; Environmental Monitoring; Environmental science; Enzymes; Fourier transforms; Hydrogen-Ion Concentration; Immunoassay; Immunoenzyme Techniques; In-line Multiplexed Biosensing; Ions; Kinetics; Luminescent Measurements - methods; Marine Toxins; Metal ions; Microcystins; Microcystins - chemistry; Peptides; Physicochemical properties; Polyclonal antibodies; Proteins; Sensitivity and Specificity; Sorption; Studies; Time Factors; Toxins; Waste Water Technology; Water Management; Water Pollution Control; Water quality; Zinc
• ISSN: 0944-1344; 1614-7499
• DOI: 10.1007/s11356-015-5758-y

A highly sensitive, specific, simple, and rapid chemiluminescence enzyme immunoassay (CLEIA) was developed for the determination of microcystin-LR (MC-LR) by using strategies for oriented immobilization of functionally intact polyclonal antibodies on chitosan surface. Several physicochemical parameters such as metal ion adsorption, hexahistidine-tagged Protein G sorption, the dilution ratio polyclonal antibody concentration, and peroxidase-labeled MC-LR concentration were studied and optimized. The sorption in batch system of G-histidine and G-proteins was studied on a novel sorbent consisting of chitosan/divalent metal ions. Transition metals as Ni ++ and Zn ++ were immobilized through interaction with –NH 2 groups of chitosan in order to supply a material capable to efficiently remove the proteins from aqueous solutions. The maximum uptake of divalent metals onto the chitosan material was found to be 230 mg g −1 for Zn ++ and 62 mg g −1 for Ni ++ . Experimental data were evaluated using the Langmuir and Freundlich models; the results were well fitted with the Langmuir model; chitosan/Ni ++ foam was found to be the best sorbent for G-protein, maximum sorption capacity obtained was 17 mg g −1 , and chitosan/Zn ++ was found to be the best for G-histidine with a maximum sorption capacity of 44 mg g −1 . Kinetic data was evaluated with pseudo-first- and pseudo-second-order models; the sorption kinetics were in all cases better represented by a pseudo-second-order model. Under optimum conditions, the calibration curve obtained for MC-LR gave detection limits of 0.5 ± 0.06 μg L −1 , the 50 % inhibition concentration (IC50) was 2.75 ± 0.03 μg L −1 , and the quantitative detection range was 0.5–25 μg L −1 . The limit of detection (LOD) attained from the calibration curves and the results obtained demonstrate the potential use of CLEIA with chitosan support as a screening tool for the analysis of pollutants in environmental samples.