Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1994-12-27
1998-05-26
Chin, Christopher L.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
436525, 436501, 436151, 436806, 422 8201, 422 8202, 422 8203, 204400, 204403, 204416, 204418, 204194, 204415, 4352871, 4352872, G01N 3353, G01N 33553, G01N 2700, G01N 2726
Patent
active
057563551
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to novel membrane sensors containing an array of self assembling lipid molecules and to the use of such sensors as biosensors.
2. Description of the Related Art
A biosensor comprises a biologically sensitive material in intimate contact with a suitable electronic device, the transducer, which converts a biochemical signal generated by the interaction between the biologically sensitive material and the surrounding fluid medium, the analyte, into quantifiable or processable electronic or optical information. A wide variety of biosensitive materials have been proposed for use in biosensors of various kinds. Enzymes, antibodies, antigens, receptor proteins, whole mammalian cells or tissues have all been suggested as being potentially useful. However in order to incorporate these materials into a sensor it is necessary to immobilise them in a way which will allow them to interact with the analyte and generate a suitable signal.
One class of biosensor is based on bilayer lipid membranes (BLMs). For a general review, see H. T. Tien et al. in "Molecular Electronics: Biosensors and Biocomputers", ed. F. T. Hong, Plenum Press, New York (1989) at pages 259-268. A sub-class of BLM biosensors mentioned therein is those which one face of the bilayer is anchored to a support while the other is in contact with a solution of analyte. The lipid bilayer is self-assembling, as the lipid molecules employed arrange themselves perpendicularly to the support in the manner: phosphate or acetate. The lipid bilayer contains a small number of biosensitive molecules. By way of illustration, PCT Application Publication No. WO 89/01159 (CSIRO) discloses BLMs containing a reagent providing an ion-channel, e.g. the peptide gramicidin. The FAb fragment of a divalent monoclonal antibody to human chorionic gonadotrophin (hCG) is present in the aqueous solution. The antibody blocks the ion-channel, but the block is partially released when hCG analyte is introduced into the aqueous solution, as demonstrated by a change in impedance. In these BLMs, the first layer is of dodecanethiol, the second an acetate lipid or phospholipid and support is a palladium-coated glass electrode. The thiol group provides bonding to the palladium surface. PCT Application Publication No. WO 90/02327 (Australian Membrane and Biotechnology Research Insitute) relates to detailed modifications of the earlier work. It mentions that if the lipid is directly coating a metal surface such as palladium, it would be necessary to substitute a thio residue for the phospholipid head group.
European Patent Application Publication EP-441 120 A (Yeda) points out the desirability of providing a water layer between the support and the BLM, by joining the phospholipid to the support by hydrophilic spacer arms. For example, the phospholipid with the spacer may be of formula (1): of oxyethylene groups is from 7 to 25. The thiol group bonds the phospholipid to the support. This mode of attachment allows the use of mixed lipids in the composition of the lipid layers, where the minor component consists of the phospholipid with the spacer that is used for attachment to the support, while the bulk of the phospholipids define the bilayer structure (page 4 lines 34-36, FIG. 2). In other words, the bilayer is attached to a recording electrode by a few bridging, anchoring molecules, leaving a gap between the lower face of the bilayer and the upper face of the electrode, which is filled by the aqeuous medium. The BLMs are doped with "ion channels" which are normally closed, but are opened in the presence of an effector molecule. Thus the BLMs may contain an acetyl choline receptor which is alleged to give an increase in conductivity when acetyl choline is added, as analyte, to the medium surrounding the upper surface of the bilayer.
H. Lang et al. have presented a poster at the 5th International Conference on Langmuir-Blodgett Films, Paris, 26-30th August 1991 which describes BLMs in which the first layer is anchored to
REFERENCES:
patent: 5192507 (1993-03-01), Taylor et al.
Bain et al., "Modeling Organic Surfaces with Self-Assembled Monolayers", Angew. Chem. Int. Ed. Engl., 28(4), pp. 506-512 (1989).
Lang et al., "Self-Assembly of Thiolipid Molecular Layers On Gold Surfaces: Optical and Electrical Characterization", Thin Solid Films, 210/211, pp. 818-821 (1992).
Markowitz et al., "Self-Assembling Properties of 1,2-Diacyl-sn-glycero-3-phosphohydroxyethanol: A Headgroup-Modified Diacetylenic Phospholipid", Langmuir, 7(1), pp. 16-18 (1991).
Swaney, John B. Mechanism of Protein-Lipid Interaction, The Journal of Biological Chemistry, vol. 255, No. 18, pp. 8791-8797, 1980.
Williams, Brian Wesley Effect of Proteins on Fluorophore Lifetime Heterogeneity in Lipid Bilayers, vol. 29, pp. 3248-3255, 1989.
Koenig Bernd
Lang Holger
Vogel Horst
Chin Christopher L.
Ecole Polytechnique Federale de Lausanne
Nguyen Bao-Thuy L.
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