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Environmental Science and Pollution Research

, Volume 24, Issue 1, pp 15–24 | Cite as

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

  • Hary Demey
  • Scherrine A. Tria
  • Romain Soleri
  • Anthony Guiseppi-Elie
  • Ingrid Bazin
In-line Multiplexed Biosensing

Abstract

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 –NH2 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.

Keywords

Chitosan Microcystin-LR Antibody orientation Protein G Affinity tag 

Notes

Acknowledgments

AGE acknowledges receipt of a visiting professorship from EMA. This work was financially supported by the French National Research Agency via funding for the COMBITOX project (ANR-11-ECOT-009-04)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Hary Demey
    • 1
  • Scherrine A. Tria
    • 2
  • Romain Soleri
    • 2
  • Anthony Guiseppi-Elie
    • 3
  • Ingrid Bazin
    • 2
  1. 1.École des Mines d’Alès, Centre des Matériaux des Mines d’AlèsAlès CEDEXFrance
  2. 2.École des Mines d’Alès, Laboratoire de Génie de L’Environnement IndustrielAlès CEDEXFrance
  3. 3.Department of Biomedical Engineering, The Dwight Look College of EngineeringTexas A&M UniversityCollege StationUSA

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