| 摘要 |
The present invention is related to a method for a quantification of a target compound (1) (being a polynucleotide or a protein) present in a sample solution, comprising the steps of:
a) Possibly diluting the target compound (1) by a known dilution factor,
b) possibly labeling the target compound (1),
c) putting into contact the target compound (1) with a capture probe (2), in order to allow a specific binding between the said target compound (1) and the said capture probe (2), the said capture probe (2) being fixed upon a surface of a solid support (3) according to an array comprising a density of at least 10 discrete regions per cm 2 , each of said discrete regions (A) being fixed with a species of capture probe (2) capable of specifically binding to a target compound (1), wherein at least 4 additional discrete regions (B) of the solid support (3) surface are printed with solutions containing known different and increased concentrations of a (labeled) detection molecule (4, 4', 4", 4"'), said detection molecule being from the same family, either a polynucleotide or a protein, as the capture probes (2),wherein each of said discrete regions (A,B) have a diameter comprised between 50 and 500 µm,
d) detecting by the same detection means signals resulting from the binding between the target compound (1) and its corresponding capture probe (2) and resulting from the printed detection molecule (4, 4', 4", 4"') in the different discrete regions (A, B),
e) obtaining a detection curve (C) of the detected signals of said detection molecule (4, 4', 4", 4"') in the at least 4 additional discrete regions (B) of the solid support (3) surface as a function of their corresponding printed detection solution concentrations in arbitrary units (AU),
f) converting the signal obtained from the target compound (1) bound to a specific capture probe (2) into a concentration value presented in arbitrary units (AU) by a conversion step upon the detection curve (C) of the detected signal, and
g) quantifying the target compound by converting said concentration value into a target amount using a target concentration curve.
|
