Elongated membrane flow-through diagnostic device and method
First Claim
1. A storage and reaction apparatus for use in assays for determining bindable target substances comprising:
- (a) a liquid-permeable, porous, elongate reaction membrane strip comprising an upper and lower surface, at least said upper surface comprising immobilized proteins capable of specifically binding to said target substances, said proteins being positioned on said strip so as to correspond to electrophoretically resolved proteins which have been transferred to said strip and which are capable of specifically binding to said target substances,(b) a body of absorbent material capable of absorbing liquid, said body having a surface located adjacent to the lower surface of the reaction membrane strip, and(c) a container means for said reaction membrane strip and said absorbent material comprising a top wall defining a fluid port in the form of an elongate slot adjacent said membrane strip and further comprising a fluid seal means peripherally disposed at said fluid port defining a seal between said container top wall and said reaction membrane strip.
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Abstract
An apparatus is provided for use in assays for the detection of a bindable target substance in a liquid sample. The apparatus comprises an elongate reaction membrane strip which is porous and liquid-permeable, the upper surface of the membrane strip having receptors thereon capable of binding to the target substance; an absorbent material located adjacent to the lower surface of the reaction membrane strip; and a container means for the reaction membrane strip and absorbent material, including a top wall defining a fluid port in the form of an elongate slot adjacent the membrane strip and including a fluid seal means disposed at the periphery of the fluid port defining a seal between the container top wall and reaction membrane strip. In one embodiment, the receptors on the membrane strip correspond to Western blot proteins. An assay for the detection of bindable target antibody in a liquid sample employing the apparatus is also described.
78 Citations
23 Claims
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1. A storage and reaction apparatus for use in assays for determining bindable target substances comprising:
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(a) a liquid-permeable, porous, elongate reaction membrane strip comprising an upper and lower surface, at least said upper surface comprising immobilized proteins capable of specifically binding to said target substances, said proteins being positioned on said strip so as to correspond to electrophoretically resolved proteins which have been transferred to said strip and which are capable of specifically binding to said target substances, (b) a body of absorbent material capable of absorbing liquid, said body having a surface located adjacent to the lower surface of the reaction membrane strip, and (c) a container means for said reaction membrane strip and said absorbent material comprising a top wall defining a fluid port in the form of an elongate slot adjacent said membrane strip and further comprising a fluid seal means peripherally disposed at said fluid port defining a seal between said container top wall and said reaction membrane strip. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A storage and reaction apparatus for use in assays for determining a bindable target substance in a liquid sample suspected of containing said target substance, comprising:
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(a) a liquid-permeable, porous, elongate reaction membrane strip comprising an upper surface, a lower surface, and side walls, at least said upper surface comprising an immobilized receptor capable of specifically binding to said target substance, (b) a body of absorbent material capable of absorbing liquid, said body having a surface located adjacent to the lower surface of the reaction membrane strip, and (c) a container means for said reaction membrane strip and said absorbent material comprising a top wall defining an elongate slot fluid port adjacent said reaction membrane strip and further comprising a fluid seal peripherally disposed at said fluid port defining a seal between said container top wall and said reaction membrane strip, said fluid seal comprises a continuous rim projecting from said container top wall towards said reaction membrane strip, said rim and said reaction membrane strip being in contact under sufficient compression to prevent leakage between said rim and said reaction membrane strip, said rim having an outer wall projecting to an area of said reaction membrane strip interior of said side walls thereby forming an open area on the top of the strip outside the rim and free of contact with the rim.
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10. An assay for determining bindable target antibodies in a liquid sample suspected of containing said antibodies, employing as an assay device an apparatus comprising:
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(a) a liquid-permeable, porous, elongate reaction membrane strip comprising an upper and lower surface, at least said upper surface comprising immobilized proteins capable of specifically binding to said target antibodies, said proteins being positioned on said strip so as to correspond to electrophoretically resolved proteins which have been transferred to said strip and which are capable of specifically binding to said target antibodies, (b) a body of absorbent material capable of absorbing liquid, said body having a surface located adjacent to the lower surface of the reaction membrane strip, (c) a container means for said reaction membrane strip and said absorbent material comprising a top wall defining a fluid port in the form of an elongate slot adjacent said reaction membrane strip and further comprising a fluid seal means peripherally disposed at said fluid port defining a seal between said container top wall and said reaction membrane strip, said assay comprising applying said liquid sample to the upper surface of said permeable reaction membrane strip whereby said antibodies, if present in said liquid sample, specifically bind to said immobilized proteins and unbound sample permeates through said fluid port and the membrane lower surface into said absorbent body;
applying a visible labelled substance capable of directly or indirectly specifically binding to said antibodies to the upper surface of said permeable reaction membrane strip whereby said visible labelled substance specifically binds any bound target antibodies and unbound visible labelled substance permeates through said fluid port and the membrane lower surface into said absorbent body; and
, thereafter determining any visible labelled substance specifically bound to said upper surface as an indication of the presence or amount of said target antibodies in said liquid sample. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. An assay for determining bindable target antibody in a liquid sample suspected of containing said antibody, employing as an assay device an apparatus comprising a liquid-permeable, porous, elongate reaction membrane strip comprising an upper and lower surface, at least said upper surface comprising immobilized protein antigen capable of specifically binding to said target antibody, a body of absorbent material capable of absorbing said sample, said body comprising a surface located adjacent to the lower surface of the reaction membrane strip, and a container means for said reaction membrane strip and said absorbent material comprising a top wall defining a port adjacent said reaction membrane strip and further comprising a fluid seal means peripherally disposed at said fluid port defining a seal between said container top wall and said reaction membrane strip, said assay comprising:
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(a) applying a blocking solution to said reaction membrane strip, said blocking solution comprising between about 1% and about 2% polyoxyethylene sorbitan monolaurate surfactant and between about 0.5% and about 1.5% immunologically non-reactive protein, (b) applying said liquid sample to the upper surface of said reaction membrane strip whereby said antibody, if present in said liquid sample, specifically binds to said immobilized proteins and unbound sample permeates through said fluid port and the membrane lower surface into said absorbent body;
applying a visible labelled substance capable of directly or indirectly specifically binding to said antibodies to the upper surface of said permeable reaction membrane strip whereby said visible labelled substance specifically binds any bound target antibody and unbound visible labelled substance permeates through said fluid port and the membrane lower surface into said absorbent body; and
,(c) thereafter determining any visible labelled substance specifically bound to said upper surface as an indication of the presence or amount of said target antibody in said liquid sample. - View Dependent Claims (22, 23)
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Specification