Immunoassay Product and Process

Abstract

A rapid, efficient and convenient method to detect one or more biological entities on a blotting membrane is provided. The detection can relate to the position, nature or amount of the biological substance on one or more membranes. The invention method involves a pressure assisted regiment (such as vacuum or positive gas pressure) for the supply and removal of reagents and permits washing of the contaminants from substances embedded in the membrane to be detected using very low volumes of liquid. This method enables completion of the blocking, washing and antibody binding steps in about 30 minutes without comprising blot quality. In another aspect, the invention is directed to an apparatus useful in conducting the method of the invention. The device is comprised of several layers including a porous support layer below the blotting membrane(s), a flow distributor above the blotting membrane(s) and a well on the flow distributor to contain the liquid to the desired area and to allow for lower starting volumes of such liquid. Preferably, the flow distributor is a non-binding or low binding hydrophilic porous membrane such as a 0.22 micron membrane.

What we claim: 
     
         1 . A device for conducting immunoassays comprising a porous support, a flow distributor placed on top of the porous support and one or more reagent wells mounted on top of the flow distributor. 
     
     
         2 . A device for conducting vacuum assisted immunoassays comprising a vacuum manifold, a porous support, at flow distributor placed on top of the porous support and one or more reagent wells mounted on top of the flow distributor. 
     
     
         3 . A device for conducting vacuum assist immunoassays comprising a porous support, one or more membranes containing one or more biological entities to be assayed, the one or more membranes being mounted on top of the porous support, a flow distributor on top of the one or more membranes and one or more reagent wells mounted on top of the flow distributor. 
     
     
         4 . A device for conducting pressure assisted immunoassays comprising a collection manifold, a porous support, one or more membranes containing one or more biological entities to be assayed, the one or more membranes being located on top of the porous support, a flow distributor placed on top of the one or more membranes, one or more reagent wells mounted on top of the flow distributor, a pressure cap removably sealed on top of the one or more reagent wells, the cap having an inlet to its interior, the inlet being connected to a source of positive gas pressure. 
     
     
         5 . The device of  claim 1  wherein the flow distributor and the one or more reagent wells are an integral unit. 
     
     
         6 . The device of  claim 1  wherein the flow distributor is a membrane and the flow distributor and one or more reagent wells are an integral unit. 
     
     
         7 . The process of  claim 7  further comprising a step (d) of detecting the one or more biological entities. 
     
     
         8 . The process of  claim 7  wherein in step (b) the reagent is an antibody and is allowed to incubate on/in the one or more blotting membranes without applying the vacuum for a set period of time. 
     
     
         9 . The device of  claim 1  further comprising a collection tray below the holder for the recovery of reagents. 
     
     
         10 . The device of  claim 1  further comprising a collection tray below the holder for the recovery of reagents and wherein the tray is subdivided into two or more separate subtrays. 
     
     
         11 . The device of  claim 1  wherein each well has a separate flow distributor.