TY - JOUR
T1 - Sorption of his-tagged Protein G and Protein G onto chitosan/divalent metal ion sorbent used for detection of microcystin-LR
AU - Demey, Hary
AU - Tria, Scherrine A.
AU - Soleri, Romain
AU - Guiseppi-Elie, Anthony
AU - Bazin, Ingrid
N1 - Funding Information:
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)
Publisher Copyright:
© 2015, Springer-Verlag Berlin Heidelberg.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - 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.
AB - 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.
KW - Affinity tag
KW - Antibody orientation
KW - Chitosan
KW - Microcystin-LR
KW - Protein G
UR - http://www.scopus.com/inward/record.url?scp=84949817234&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84949817234&partnerID=8YFLogxK
U2 - 10.1007/s11356-015-5758-y
DO - 10.1007/s11356-015-5758-y
M3 - Article
C2 - 26667644
AN - SCOPUS:84949817234
SN - 0944-1344
VL - 24
SP - 15
EP - 24
JO - Environmental Science and Pollution Research
JF - Environmental Science and Pollution Research
IS - 1
ER -