Synthetic resins or natural rubbers -- part of the class 520 ser – Synthetic resins – Mixing of two or more solid polymers; mixing of solid...
Reexamination Certificate
1999-03-12
2001-01-23
Nutter, Nathan M. (Department: 1711)
Synthetic resins or natural rubbers -- part of the class 520 ser
Synthetic resins
Mixing of two or more solid polymers; mixing of solid...
C521S099000, C210S660000, C210S661000, C210S663000, C252S408100, C436S008000, C436S163000
Reexamination Certificate
active
06177513
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a method for evaluating artificial receptors, more particularly an evaluation method for use in development of artificial receptors, which have been recently growing in use as separating agents for chemicals, sensor materials for chemicals, reagents for immunoassays and the like.
2. Description of Related Art
In living bodies, a variety of functions are expressed by proper recognition of a huge amount of chemical substances. For example, an enzyme effectively catalyzes a chemical reaction in a living body by recognizing a specific substance. Also various kinds of receptors existing in cell membranes bind selectively to specific substances and transfer extracellular information received from the specific substances into cells. The cells in turn exercise functions required in a living body such as suppression, regulation and the like on the basis of the extracellular information.
Artificial receptors are those artificially designed and synthesized to imitate such receptors and exhibit good molecular recognition functions. For designing and synthesizing such artificial receptors, there are proposed several methods, among which “Molecular Imprinting method by Wuff, G.,
Angew.Chem.,Int.Ed.Engl
.,34,1812-1832(1995) is said to be particularly excellent and preferable.
According to the Molecular Imprinting method (hereinafter referred to as MI method), an artificial polymeric receptor can be prepared by polymerizing a polymerizable organic compound (a functional monomer) which interacts with a template substance (also referred to as a key molecule, imprint molecule or template molecule) and a crosslinker(i.e., crosslinking monomer) in the presence of the template substance and then releasing and removing the template substance from the resultant polymer. This polymeric receptor is a polymer characterized by having a cavity fit to the molecular morphology and size of the template substance. In addition to that, the polymer obtained by the MI method (hereinafter referred to as a molecularly imprinted polymer) is physically and chemically stable and has an excellent storage endurance.
Published Japanese Patent Application No. HEI 8(1996)-506320 (corresponding to the International Patent Application Publication No. WO94/11403) discloses the use of a molecularly imprinted polymer (artificial antibody) for detecting an antigen in immunoassay.
Further, Japanese Unexamined Patent Publication No. HEI 8(1996)-15160 discloses a biomimetic sensor using a thin molecularly imprinted polymer.
With a view to obtaining a suitable molecularly imprinted polymer, however, the MI method has several problems. First, it is necessary to study suitable polymerization conditions about combination and proportion of components, i.e., the functional monomer and crosslinker, with respect to the template substance as well as a solvent to be used for polymerization. Moreover, in order to evaluate the obtained molecularly imprinted polymer, the polymer must be crushed into pieces, packed in a column and evaluated about to what extent the polymer can recognize the template substance. This whole evaluation process consumes considerable time and labor.
A conventional in situ method by J. Matsui, T. Kato, T. Takeuchi, M. Suzuki, K. Yokoyama, E. Tamiya, and I. Karube,
Anal.Chem
., 65, 2223(1993), which was developed for reducing such time and labor, discloses a method for preparing a molecularly imprinted polymer directly in a column for HPLC by feeding a mixture of a template substance, functional monomer(s), a crosslinker, a solvent, an initiator for polymerization and the like in the column and polymerizing in situ. According to this method, the ability of the molecularly imprinted polymer to recognize the template molecule can be evaluated only by connecting the resulting column to a pump for HPLC and subjecting to HPLC analysis. However, this method is not a satisfactory one because it still needs much time and work.
SUMMARY OF THE INVENTION
The present inventors have found a new in situ method for evaluating an artificial receptor. Accordingly, the present invention provides a method for evaluating an artificial receptor, which includes subjecting a functional monomer and a crosslinker in the presence of a template substance(i.e., a substance to be recognized) in a vessel into polymerization, thereby forming a resultant polymer with the template substance usually in the form of a thin film; dissociating the template substance from the polymer and removing it from the vessel by washing the inside of the vessel, thereby to prepare an artificial receptor; feeding a solution of the same or different template substance into the vessel to associate the template substance with the artificial receptor; and then measuring an amount of the template substance remaining in the vessel and evaluating the artificial receptor on the basis of a determination result.
In another aspect, the present invention provides an artificial receptor obtained by the above-mentioned method of the invention.
In a further aspect, it provides a method for quantitating a template substance comprising the steps of: subjecting a functional monomer and a crosslinker in the presence of a template substance in a vessel into polymerization, thereby forming a resultant polymer with the template substance usually in the form of a thin film; dissociating the template substance from the polymer and removing it from the vessel by washing an inside of the vessel, thereby to prepare an artificial receptor; feeding a sample of a template substance, which may be the same or different with the above template substance, in an unknown concentration into the vessel to associate the template substance with the artificial receptor, separating the associated product; and measuring an optical parameter of the template substance in the artificial receptor.
REFERENCES:
patent: 8-15160 (1996-01-01), None
patent: 8-50632 (1996-07-01), None
Gunter Wulff, “Molecular Imprinting in Cross-Linked Materials with the Aid of Molecular Templates”, pp. 1812-1832; Angew. Chem. Int Ed. Engl., 34 (1995).
Jun Matsui et al., Analytical Chemistry, 67 (1995), “A Molecularly Imprinted Synthetic Polymer Receptor Selective for Atrazine”, pp. 4404-4408.
Jun Matsui et al., Chemistry Letters, (1995), “Atrazine-selective Polymer Prepared by Molecular Imprinting Technique”, p. 489.
Jun Matsui et al., Analytical Chemistry, 65 (1993), “Molecular Recognition in Continuous Polymer Rods Prepared by a Molecular Imprinting Technique”, pp. 2223-2224.
Fukuma Daigo
Matsui Jun
Mizukami Toshihiro
Takeuchi Toshifumi
Birch & Stewart Kolasch & Birch, LLP
Nutter Nathan M.
Toshifumi Takeuchi
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