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Journal of Automatic Chemistry
Volume 12 (1990), Issue 2, Pages 53-59
http://dx.doi.org/10.1155/S1463924690000074

Determination of specific proteins by the FIA principle

1Clinical Chemistry Department, Copenhagen County Hospital in Herlev, Herlev, DK 2730, Denmark
2Immuno Chemistry Department, Novo-Nordisk A/S, Sauntesvej 13, Gentofte, DK 2820, Denmark

Copyright © 1990 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The following analytes have been investigated: urine albumin (u-albumin), plasma-transferrin (p-transferrin), p-haptoglobin, p-IgG, p-IgA, p-IgM, and p-orosomucoid. An unmodified commercial analytical system FIA Star (Tecator) with a two-channel injector (40 μl) was used. The prediluted plasma samples and antibodies are allowed to react for 33 s before the change in turbidity is measured as a Peak maximum at 405 nm. The optimal concentrations of calibrators and antibodies have been determined to secure antibody excess. Response time (i.e. delay between aspiration of a sample and presentation of the result in absorption units) is 75 s. Automatic print-out of the absorbance profile and movement of the sample rack further accounted for 21 s per sample, so the throughput is reduced to 75 determinations per 2 h. Results are available within an hour, compared to two-12 days with the present methods (electroimmunoassays). Parallel analyses with established methods/analysers show excellent agreement for u-albumin, p-transferrin and p-haptoglobin. For p-IgG, p-IgA and p-IgM the reaction time of 33 s is insufficient because their relative molecular masses (i.e. the size of the molecules) are so high, 150.000-971.000. Five minutes is a more adequate reaction time, which makes a serial analyser such as FIA Star unsuitable for larger workloads of samples of immunoglobulins. The plasma concentration of Orosomucoid is low, resulting in high sample blanks. It is therefore recommended that the reaction is followed kinetically if a serial analyser is used.