Patent ID: 7166473

Claim:
A method of producing an algorithm for determining the content of a transferrin variant or combination of transferrin variants, which may be used as a diagnostic tool for alcohol consumption, in a sample of body fluid, said method comprising: (a) obtaining at least two solutions, each having known contents of asialo- (A1, A2, A3, etc.) and disialo-transferrins (D1, D2, D3, etc.); (b) subjecting each of said solutions to a separation method to separate fractions substantially free from tri- and higher sialylated transferrins; (c) determining the total transferrin variant content (T1, T2, T3, etc.) of said fractions; (d) determining the correlation between the content of asialo- and disialo-transferrins or between the content of asialo-, monosialo, and disialo-transferrins in the sample; (e) producing a calibration curve based on the determined correlation between the content of asialo- and disialo-transferrins or the content of asialo-, monosialo, and disialo-transferrins in the sample; (f) using the calibration curve to produce an algorithm that relates the content of asialo- and disialo-transferrins or the content of asialo-, monosialo, and disialo-transferrins in the sample to the content or amount of one or more transferrin variants in the sample; and (g) determining the content of a transferrin variant or combination of transferrin variants in the sample.