The gramicidin-based biosensor: a functioning nano-machine.

Biosensors combine a biological recognition mechanism with a physical transduction technique. In nature, the transduction mechanism for high sensitivity molecular detection is the modulation of the cell membrane ionic conductivity through specific ligand-receptor binding-induced switching of ion channels. This effects an inherent signal amplification of six to eight orders of magnitude, corresponding to the total ion flow arising from the single channel gating event. Here we describe the first reduction of this principle to a practical sensing device, which is a planar impedance element composed of a macroscopically supported synthetic bilayer membrane incorporating gramicidin ion channels. The membrane and an ionic reservoir are covalently attached to an evaporated gold surface. The channels have specific receptor groups attached (usually antibodies) that permit switching of gramicidin channels by analyte binding to the receptors. The device may then be made specific for the detection of a wide range of analytes, including proteins, drugs, hormones, antibodies, DNA, etc., currently in the 10(-7)-10(-13) M range. It also lends itself readily to microelectronic fabrication and signal transduction. By adjusting the surface density of the receptors/channel components during fabrication, the optimum sensitivity range of the device may be tuned over several orders of magnitude.

A biosensor that uses ion-channel switches.

Biosensors are molecular sensors that combine a biological recognition mechanism with a physical transduction technique. They provide a new class of inexpensive, portable instrument that permit sophisticated analytical measurements to be undertaken rapidly at decentralized locations. However, the adoption of biosensors for practical applications other than the measurement of blood glucose is currently limited by the expense, insensitivity and inflexibility of the available transduction methods. Here we describe the development of a biosensing technique in which the conductance of a population of molecular ion channels is switched by the recognition event. The approach mimics biological sensory functions and can be used with most types of receptor, including antibodies and nucleotides. The technique is very flexible and even in its simplest form it is sensitive to picomolar concentrations of proteins. The sensor is essentially an impedance element whose dimensions can readily be reduced to become an integral component of a microelectronic circuit. It may be used in a wide range of applications and in complex media, including blood. These uses might include cell typing, the detection of large proteins, viruses, antibodies, DNA, electrolytes, drugs, pesticides and other low-molecular-weight compounds.

Chemical shift anisotropies obtained from aligned egg yolk phosphatidylcholine by solid-state 13C nuclear magnetic resonance.

Natural abundance solid-state 13C NMR spectra were obtained from orientated egg yolk phosphatidylcholine multilayers in which peaks from the different types of carbon in the lipid were resolved. The residual chemical shift anisotropy of the choline, glycerol, and olefinic carbons, as well as the carbonyl and acyl chain methylene carbons, were estimated. This information provided the basis for a qualitative description of the order and conformation of egg yolk phosphatidylcholine in the L alpha phase. The results suggested the gauche conformation for the C alpha-C beta bond in the choline moiety, a constrained glycerol region, a magic angle orientation for the sn-2 carbonyl, and a preferred orientation close to the bilayer normal for the plane of the sn-1 carbonyl bond and acyl chain C = C bond. The orientations of the carbon nuclei are in accord with the molecular conformation derived from previous 2H, 31P, and 13C NMR studies.