ABSTRACT
In situ steady-state fluorescence (SSF) measurement technique was applied to investigation of pyrene labeled Poly(vinyl alcohol) (PVA-Py) molecules diffusion in and out of agarose gels. Gel samples with four different concentration of agarose were prepared. PVA-Py was synthesized by "click" chemistry method and dissolved in water to use in diffusion experiments. The results were analyzed by using Fickian type diffusion model, and it was found that sorption and desorption processes of PVA-Py molecules in and out of agarose gel have two distinct regions for short and long diffusion times. Sorption and desorption coefficients were measured and it was seen that the diffusion rates were much larger at short times and at lower agarose concentrations.
Subject(s)
Polyvinyl Alcohol/chemistry , Pyrenes/chemistry , Sepharose/chemistry , Diffusion , Gels , Spectrometry, FluorescenceABSTRACT
Poly(vinyl alcohol)-pyrene-anti-metadherin (PVA-Py-(Anti-MTDH)), a novel antibody based water soluble probe containing both fluorescent and target sites in the structure for in vitro imaging of breast cancer cells is reported here. Since breast cancer cells have an excess of MDTH protein expressed on the surface, a PVA-Py prepared by "Click chemistry" approach is targeted by Anti-MTDH antibody and applied to the MCF-7 cell line. After characterization, the designed architecture was evaluated in terms of cell incorporation efficiency and compared with a non-targeted structure (PVA-Py). Atomic force microscopy (AFM) and fluorescence microscopy images of cells after incubation of the probe molecules were also obtained to monitor the interaction of the probes with the cancerous cells.
Subject(s)
Breast Neoplasms/pathology , Cell Adhesion Molecules/immunology , Diagnostic Imaging/methods , Immunoconjugates , Antibodies, Neoplasm/chemistry , Breast Neoplasms/diagnosis , Cell Adhesion Molecules/analysis , Cell Line, Tumor , Female , Fluorescent Dyes/chemistry , Humans , Immunoconjugates/chemistry , Immunoconjugates/immunology , Membrane Proteins , Neoplasm Proteins/analysis , Neoplasm Proteins/immunology , Polyvinyl Alcohol/chemistry , Pyrenes/chemistry , RNA-Binding ProteinsABSTRACT
Poly(vinyl alcohol)-pyrene-glucose oxidase (PVA-Py-GOx), a water-soluble polymer possessing both fluorescent and oxidant sites in the structure, is synthesized by "click" chemistry and modification processes and characterized. The morphology of PVA-Py-GOx was characterized with atomic force microscopy (AFM), and a heterogeneous morphology due to the incorporation of GOx was observed. The capability of PVA-Py-GOx to act as a bioprobe for fluorescence sensing of glucose is examined. The postulated fluorescence mechanism for glucose analysis is based on the consumption of glucose by dissolved oxygen and GOx present in the structure. Thus, the fluorescence intensity of pyrene groups of the probe increases by the elimination of fluorescence quenching by oxygen. Glucose concentration was analyzed quantitatively from 0.25 to 3.0 mM by the fluorescence measurement. The effect of pH and amount of PVA-Py-GOx was also studied. The proposed system was applied to analyze glucose in real samples and compared with those obtained from commercial kits.
