title: Development of a label-free SPR sensor for detection of matrixmetalloproteinase-9 by antibody immobilization on carboxymethyldextran chip.
abstract: Surface plasmon resonance (SPR) immunosensor has been widely utilized for monitoring antigen-antibody interactions. The sensor measures changes of refractive index upon binding of analyte molecules to specific ligand immobilized on the sensor chip. This effort reports development of SPR immunosensor for real-time and label-free detection of recombinant human matrix metalloproteinases-9 (MMP-9), which has been associated with malignant tumor progression and metastasis by matrix degradation. MMP-9 was expressed in Escherichia coli BL21 and purified by Ni-NTA agarose column. CMD 50 D was activated by EDC/NHS for immobilization of monoclonal anti-MMP-9. Atomic force microscopy images showed uniform distribution of anti-MMP-9 over the sensor chip. Equilibrium constant (KD), maximum binding capacity (Rmax) and ∆Gb values for interaction of MMP-9 and anti-MMP-9 were 0.4nM, 680 µRIU and -53.51kJ/mol, respectively. Concentration of MMP-9 in saliva samples was determined, with linearity in the range of 10-200ng/mL. The limit of detection was found to be 8pg/mL, being lower than most of the previously reported techniques.
authors.affiliation: Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Sara
authors.initials: S
authors.lastname: Mohseni
authors.affiliation: Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Tahereh Tohidi
authors.initials: TT
authors.lastname: Moghadam
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Bahareh
authors.initials: B
authors.lastname: Dabirmanesh
authors.affiliation: Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Safoura
authors.initials: S
authors.lastname: Jabbari
authors.affiliation: Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran; Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran. Electronic address: khajeh@modares.ac.ir.
authors.forename: Khosro
authors.initials: K
authors.lastname: Khajeh
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry; D003911:Dextrans / Q000737:chemistry*; D004867:Equipment Design; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation; D057230:Limit of Detection; D020780:Matrix Metalloproteinase 9 / Q000032:analysis*; D012031:Refractometry; D012463:Saliva / Q000201:enzymology*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
chemical_list: D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D003911:Dextrans; C014392:carboxymethyl dextran; D020780:Matrix Metalloproteinase 9
keywords: Chronic periodontitis; Matrixmetalloproteinase-9; Refractive index; Sensor chip; Surface plasmon resonance