Electrochemical detection of endotoxin using recombinant factor C zymogen

We have developed a zymogen-based electrochemical sensor. Zymogen is an inactive enzyme precursor (proenzyme) and it is necessary to transform it biochemically (e.g., by hydrolysis and conformational change) to make it an active enzyme. In this study, we demonstrated the detection of endotoxin by using recombinant Factor C (rFC), which is a protease zymogen activated by endotoxin binding. The activated rFC hydrolyzes a synthetic substrate of Boc-Val-Pro-Arg-p-nitroanilnide to generate an electrochemical active compound, p-nitroaniline (pNA). The liberated pNA was detected by differential pulse voltammetry at –0.75 V. By using this electrochemical process, 5000 endotoxin units (EU) L^?1 and 1000 EU L^?1 were detected in a Tris-Ac buffer with a pH of 7.5 at 37 °C for reaction times of 1 h and 3 h, respectively. The concept of zymogen-based electrochemical sensors is expected to lead to the development of new biosensors.     

Highly selective photoinduced perfluoroalkylation of vinylsilanes and its application to synthesis of water-shedding polysilanes

The functionalization of polysilanes is an important subject in materials science because functionalized polysilanes are expected to exhibit potentially innovative properties. This research aims at the addition of a water-shedding property to polysilanes by introducing perfluoroalkyl groups into their skeleton. The photoinduced iodoperfluoroalkylation of various vinylsilanes takes place successfully upon irradiation with a xenon lamp: vinylmonosilanes undergo iodoperfluoroalkylation with perfluoroalkyl iodides (R_fI) regioselectively, and the corresponding perfluoroalkylated silanes are obtained in moderate to high yields. Detailed optimization of the photoinduced iodoperfluoroalkylation has been investigated to apply this method to the functionalization of polysilanes. Polysilanes having vinyl groups can be synthesized by the reductive coupling of dichlorovinylsilanes with samarium diiodide (SmI₂) and samarium metal (Sm) upon irradiation with visible light. The synthesized vinylpolysilanes and R_fI (about 1.0 mM CHCl₃ solution) are coated on a glass plate sequentially, and then the following photoirradiation with light of a wavelength over 300 nm successfully adds an excellent water-shedding property to the glass plate.     

Addressable electrode array device with IDA electrodes for high-throughput detection

An electrochemical device is proposed for high-throughput electrochemical detection that consists of 32 row and 32 column electrodes on a single glass substrate. The row and column electrodes are connected to interdigitated array (IDA) electrodes to form 1024 (32 × 32) addressable sensor points in the device. Electrochemical responses from each of the 1024 sensors were successfully acquired on the device within 1 min using redox cycling at individual IDA electrodes, which ensures application of the device to comprehensive, high-throughput electrochemical detection for enzyme-linked immunosorbent assay (ELISA), reporter gene assay for monitoring gene expressions, and DNA analysis.     

J-aggregation of a sulfonated amphiphilic porphyrin at the air-water interface as a function of pH

π-A isotherms, ellipsometric measurements, Brewster angle microscopy (BAM) and reflection spectroscopy have been utilized to characterize the films of an amphiphilic porphyrin ((OD)(3)TPPS(3)) at the air-water interface as a function of pH. This porphyrin forms stable mono-molecular layers at such interfaces, and exhibits different J-aggregation as a function of pH. The J-aggregation of (OD)(3)TPPS(3) on neutral pH subphases is notable considering that the nitrogen atoms at the central macrocycle have a pK(a)≈4.9. The type of aggregates at neutral pH is like those detected at pH<4, because the central porphyrin ring is already protonated. However at basic pH the aggregation happens without protonation of the central ring but can be instead controlled by application of the surface pressure. At the air-water interface, (OD)(3)TPPS(3) shows two bands, a red component and a blue component, which have characteristics of non-degenerate linear oscillators being perpendicularly polarized between each other. The spectral behavior observed on subphases at different pHs is qualitatively interpreted by means of exciton coupling theory, assuming that the degenerate transitions attributed to the Soret band are split. Additionally, highly oriented molecular films of these J-aggregates were deposited onto transparent quartz slides.     

Densely grafted polyisocyanides synthesized by two types of polymerization techniques

A series of novel polyisocyanide‐graft‐polystyrenes and polyisocyanide‐graft‐[polystyrene‐block‐poly(butyl acrylate)]s were synthesized through the grafting‐through and grafting‐from routes with two types of living polymerization techniques: polymerization with the Pd–Pt μ‐ethynediyl dinuclear complex as the initiator and catalyst for the polyisocyanide backbone and atom transfer radical polymerization for the grafted side chain. Through the introduction of a chiral center at the side chain of the polyisocyanide backbone, helical grafted and graft block polyisocyanides were prepared through the grafting‐from method. All of the obtained polymers exhibited polydispersities in the range of 1.07–1.41. This might have been the first time grafted polyisocyanides were prepared, especially helical grafted polyisocyanides, through the operation of two living polymerization techniques. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 1871–1880, 2003