PROCESS FOR THE QUANTITATIVE DETERMINATION OF SUBSTANCES CAPABLE OF FORMING MUTUAL SPECIFIC BONDS

摘要

1489913 Quantitative analysis MILES LABORATORIES Inc 9 Oct 1974 [11 Oct 1973] 43781/74 Heading G1B A device for use in the determination of one of a specific binding pair of substances in the form of a liquid sample comprises a column through which the liquid sample can flow from inlet to outlet and containing a matrix which is porous and insoluble in the liquid sample and which has a specific binding partner for the substance to be determined fixed thereto by a chemical bond. The selective binding pair may be an antigen and its antibody, a hapten and its antibody, an enzyme and its substrate, a hormone and its receptor or a vitamin and its receptor. The matrix may comprise a polymeric material. A suitable device is shown in the Figure and comprises body 11 sealable by friction fit cap 12 and filled with a quantity of matrix 15. A preservative mixture e.g. saline solution, or fluid sample 16 may be retained in the upper part of the body. The device may be used in a method for the quantitative determination of one of a specific binding pair of substances, which method comprises (a) contacting a matrix contained in a column with a substance to be determined and with a labelled component comprising a labelled form of one of the specific binding pair of substances, the matrix having a specific binding partner to the substance being determined fixed thereto by a chemical bond; (b) after a predetermined period of time, contacting the matrix with an eluting liquid capable of eluting from the column most of the unbound labelled component; and (c) determining the relative proportion of the labelled component which is retained in the column, this proportion being a function of the amount of the substance to be determined contacted with the matrix in step (a). The labelled component may be labelled by such means as radio-active labels and enzyme-substrate labels. Examples describe the determination of insulin, Vitamin B 12 , human placental lactogen and hepatitis B antigen.