Interfacial recognition of human prostate-specific antigen by immobilized monoclonal antibody: effects of solution conditions and surface chemistry.

The specific recognition between monoclonal antibody (anti-human prostate-specific antigen, anti-hPSA) and its antigen (human prostate-specific antigen, hPSA) has promising applications in prostate cancer diagnostics and other biosensor applications. However, because of steric constraints associated with interfacial packing and molecular orientations, the binding efficiency is often very low. In this study, spectroscopic ellipsometry and neutron reflection have been used to investigate how solution pH, salt concentration and surface chemistry affect antibody adsorption and subsequent antigen binding. The adsorbed amount of antibody was found to vary with pH and the maximum adsorption occurred between pH 5 and 6, close to the isoelectric point of the antibody. By contrast, the highest antigen binding efficiency occurred close to the neutral pH. Increasing the ionic strength reduced antibody adsorbed amount at the silica-water interface but had little effect on antigen binding. Further studies of antibody adsorption on hydrophobic C8 (octyltrimethoxysilane) surface and chemical attachment of antibody on (3-mercaptopropyl)trimethoxysilane/4-maleimidobutyric acid N-hydroxysuccinimide ester-modified surface have also been undertaken. It was found that on all surfaces studied, the antibody predominantly adopted the 'flat on' orientation, and antigen-binding capabilities were comparable. The results indicate that antibody immobilization via appropriate physical adsorption can replace elaborate interfacial molecular engineering involving complex covalent attachments.

Exploiting Chinese character models to improve speech recognition performance

The Chinese language is based on characters which are syllabic in nature. Since languages have syllabotactic rules which govern the construction of syllables and their allowed sequences, Chinese character sequence models can be used as a first level approximation of allowed syllable sequences. N-gram character sequence models were trained on 4.3 billion characters. Characters are used as a first level recognition unit with multiple pronunciations per character. For comparison the CU-HTK Mandarin word based system was used to recognize words which were then converted to character sequences. The character only system error rates for one best recognition were slightly worse than word based character recognition. However combining the two systems using log-linear combination gives better results than either system separately. An equally weighted combination gave consistent CER gains of 0.1-0.2% absolute over the word based standard system. Copyright © 2009 ISCA.

In vivo regulation of the Vi antigen in Salmonella and induction of immune responses with an in vivo-inducible promoter.

Salmonella enterica serovar Typhi, the agent of typhoid fever in humans, expresses the surface Vi polysaccharide antigen that contributes to virulence. However, Vi expression can also be detrimental to some key steps of S. Typhi infectivity, for example, invasion, and Vi is the target of protective immune responses. We used a strain of S. Typhimurium carrying the whole Salmonella pathogenicity island 7 (SPI-7) to monitor in vivo Vi expression within phagocytic cells of mice at different times after systemic infection. We also tested whether it is possible to modulate Vi expression via the use of in vivo-inducible promoters and whether this would trigger anti-Vi antibodies through the use of Vi-expressing live bacteria. Our results show that Vi expression in the liver and spleen is downregulated with the progression of infection and that the Vi-negative population of bacteria becomes prevalent by day 4 postinfection. Furthermore, we showed that replacing the natural tviA promoter with the promoter of the SPI-2 gene ssaG resulted in sustained Vi expression in the tissues. Intravenous or oral infection of mice with a strain of S. Typhimurium expressing Vi under the control of the ssaG promoter triggered detectable levels of all IgG subclasses specific for Vi. Our work highlights that Vi is downregulated in vivo and provides proof of principle that it is possible to generate a live attenuated vaccine that induces Vi-specific antibodies after single oral administration.

A surface acoustic wave-based immunosensing device using a nanocrystalline ZnO film on Si

The novelty of this study resides in the fabrication of a bio-sensing device, based on the surface acoustic wave (SAW) on a nanocrystalline ZnO film. The ZnO film was deposited using an rf magnetron sputtering at room temperature on silicon. The deposited films showed the c-axisoriented crystallite with grain size of ∼40 nm. The immunosensing device was fabricated using photolithographic protocols on the film. As a model biomolecular recognition and immunosensing, biospecific interaction between a 6-(2,4-dinitrophenyl) aminohexanoic acid (DNP) antigen and its antibody was employed, demonstrating the shifts of resonant frequencies on SAW immunosensing device. The device exhibited a linearity as a function of the antibody concentration in the range of 20∼20,000 ng/ml. © 2009 American Scientific Publishers. All rights reserved.