Concentrating zone method in heterogeneous immunoassays

C - Chemistry – Metallurgy – 12 – Q

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167/41, 167/43,

C12Q 1/70 (2006.01) G01N 33/558 (2006.01)

Patent

CA 1174595

ABSTRACT OF THE DISCLOSURE Method and apparatus are provided for for performing immunoassays employing a device comprising a relatively small test zone referred to as an immunosorbing zone, and a rela- tively large liquid absorbing zone in liquid receiving relationship with said immunosorbing zone. The immunosorbing zone includes a member of an immunological pair ("mip") - ligand and antiligand - bound to a support. A signal producing system is employed in conjunc- tion with said device having as one component a signal label bound to a mip. The signal producing system provides for production of a detectible signal in the immunosorbing zone in relation to the amount of analyte in a sample. In performing the method, the immunosorbing zone is contacted with the sample in a solution which may include one or more members of the signal producing system. The solution substantially uniformly diffuses through the immunosbosorbing zone resulting in concentration in the immunosorbing zone of the analyte and any members of the signal producing system present in the sample solution. The immunosorbing zone may then be contacted with one or more solutions to wash away any non-specific binding signal label from the immunosorbing zone and to provide any additional members of the signal producing system to the immunosorbing one for development of the detectible signal in the immunosorbing zone. The device provides that all fluids employed in the method pass through the immunosorbing zone into the liquid absorbing zone, which acts as a receptacle and driving force for drawing the fluid through the immunosorbing zone. By appropriate choice of size, conformation and material of the liquid absorbing zone, the rate and amount of fluid which passes through the immunosorbing zone at each of the stages of the method can be controlled. Thus, large volumes of liquid and large absolute amounts of reagents may be drawn through a narrow zone with the reagents at relatively low concentration as compared to the local concentration attained in the immunosorbing zone, while ensuring the sub- stantial absence of interference from non-specific binding of label and signal resulting from transient reagent.

383264

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