Patent ID: 6916666

Claim:
A lateral flow method for the determination of an analyte in a sample involving utilizing biospecific affinity reactions, and comprising the following steps: i. forming a complex in a lateral flow matrix, the complex comprising: Reactant I - - - Analyte′ - - - Reactant*, where a. Reactant* and Reactant I exhibit biospecific affinity to the analyte, b. Reactant* is analytically detectable, c. Analyte′ is the analyte or an analyte-related reactant, and subsequently ii. determining the detectable signal constituting a sample value from Reactant* in the complex, and iii. determining the amount of analyte in the sample by comparing the sample value with one or more calibrator values, each of which corresponds to a standard amount of analyte, wherein A) before determination of the calibrator value, either (i) calibrator, or (ii) a binder for the calibrator has been bound to a matrix, and when a binder for the calibrator has been bound to the matrix, calibrator is added or calibrator predeposited in the matrix is released for binding with the binder, and the matrix is insoluble in the liquid medium in which binding of Reactant* to the calibrator occurs, B) the calibrator and the analyte exhibit biospecific affinity to Reactant* by equivalent binding sites, C) two or more calibrator zones CZ comprising calibrator or binder for the calibrator are located in a single process flow stream with Reactant I in a detection zone (DZ), and D) all of the detection zones DZ are downstream of all of the calibrator zones CZ in the lateral flow matrix.