Di-tert-butylsilylene-directed alpha-selective synthesis of p-nitrophenyl T-antigen analogues.

Seven analogues of p-nitrophenyl T-antigen [Galbeta(1-->3)GalNAcalpha(1-->O)PNP] have been synthesized as potential substrates for elucidation of the substrate specificity of endo-alpha-N-acetylgalactosaminidase. These compounds, which are commercially unavailable, include: GlcNAcbeta(1-->3){GlcNAcbeta(1-->6)}GalNAcalpha(1-->O)PNP [core 4 type], GalNAcalpha(1-->3)GalNAcalpha(1-->O)PNP [core 5 type], GlcNAcbeta(1-->6)GalNAcalpha(1-->O)PNP [core 6 type], GalNAcalpha(1-->6)GalNAcalpha(1-->O)PNP [core 7 type], Galalpha(1-->3)GalNAcalpha(1-->O)PNP [core 8 type], Glcbeta(1-->3)GalNAcalpha(1-->O)PNP and GalNAcbeta(1-->3)GalNAcalpha(1-->O)PNP. The assembly of these synthetic probes was accomplished efficiently, based on di-tert-butylsilylene(DTBS)-directed alpha-galactosylation as a key reaction.

Electrochemical immunosensors for the simultaneous detection of two tumor markers.

The microfabrication of electrochemical immunosensors for the simultaneous detection of two protein analytes is described. The sensors consisted of two iridium oxide electrodes (1-mm diameter) patterned on a glass substrate. Capture antibodies were immobilized on the porous iridium oxide electrodes by covalent attachment using (3-aminopropyl)triethoxysilane and glutaraldehyde. The spatial separation of the electrodes (2.5 mm) enabled simultaneous electrochemical immunoassays to be conducted without cross-talk between the electrodes. Proteins were measured using electrochemical ELISA, and detection was achieved by electrochemically oxidizing alkaline phosphatase-generated hydroquinone. Sensors for the simultaneous detection of goat IgG and mouse IgG, and for the tumor markers CEA and AFP, were developed. The sensors had detection limits of 1, 2, 1.2, and 1 ng/mL for goat IgG, mouse IgG, CEA, and AFP, respectively.

Using antibodies to perturb the coordination sphere of a transition metal complex.

Metal ions in the active sites of many metalloenzymes exhibit distinctive spectral and chemical features which are different from those of small inorganic complexes. These features are the result of the unusual geometric and electronic constraints that are imposed on the metal ion within the protein environment. Much effort has been invested to try to mimic this feature of metalloenzymes in synthetic systems, but this remains a formidable task. Here we show that one of the key lessons learned from the science of catalytic antibodies--that binding energy can be converted into chemical energy--can be exploited to 'fine-tune' the physicochemical properties of a metal complex. We show that an antibody's binding site can reversibly perturb the coordination geometry of a metal ion, and can stabilize a high-energy coordinated species. Specifically, antibodies designed to bind the organosilicon compound 1 also bind the geometrically similar Cu(I) complex 2. However, the antibody binds a slightly compressed form of 2, which is closer in size to 1. This distortion is manifested by a spectral shift--an 'immunochromic' effect.