Liquid purification or separation – Processes – Liquid/liquid solvent or colloidal extraction or diffusing...
Patent
1985-12-03
1986-12-02
Adee, John
Liquid purification or separation
Processes
Liquid/liquid solvent or colloidal extraction or diffusing...
210656, 210927, 436161, B01H 1508
Patent
active
046263551
DESCRIPTION:
BRIEF SUMMARY
The invention is concerned with the determination of an individual person's alcohol consumption by means of quantifying isotransferrin in an isotransferrin-containing body fluid sample from the individual to be tested. This method utilizes the finding that isotransferrins of a certain pI can be correlated with an individual's alcohol comsumption; the particular names (different identities) of these isotransferrins are irrelevant and thus do not impose any restrictions on the practicability of the invention. On the basis of the correlation established between isotransferrins and alcohol consumption the invention can be a helpful tool in deciding about an adequate treatment program in each specific situation.
pH and pI values will vary with temperature and ion concentration. The pH and pI values set forth in the present specification and claims apply to a temperature of 23.degree. C. and are measured in an aqueous solution of piperazine (20 mM total) buffered with formic acid to the pH or pH=pI in question. Thus the invention is not to be construed as being restricted to the particular values set forth: It comprises also any cases involving corresponding pH and pI values at other temperatures and ion concentrations.
Two major methods have been proposed earlier for determining the content of the aforesaid isotransferrins in a serum sample. One method is based on isoelectric focusing (IEF) (Stibler, H., Borg, S. and Allgulander C. Acta Med. Scand. 206 (1979), p. 275-81); it utilizes the fact that different isotransferrins migrating in a pH gradient under the action of an electric field will cover different distances depending on the pI values of the individual iso forms, these individually different pI values in turn being due to dissimilar contents of ionizable groups. In our particular case, the difference in pI values is believed to be due inter alia to different contents of sialic acid groups; it appears that isotransferrins have been discovered which have from 0 up to 5 sialic acid groups and have isoelectric points of 6.1, 5.9, 5.7, 5.5, 5.3 and 5.1, respectively. These isotransferrins will be referred to below as being, in that same order, asialotransferrin, monosialotransferrin, disialotransferrin, trisialotransferrin, tetrasialotransferrin and pentasialotransferrin, respectively. It is possible that each of these isotransferrins will be found later on to actually consist of a plurality of individual isotransferrins; however, for the sake of simplicity they are here treated as just one substance each.
The second one of the aforesaid two methods relies on utilizing the phenomenon that different isotransferrins have different types of terminal carbohydrate groups. Thus for instance, a desialo form characteristically lacks a sialyl group on at least one carbohydrate chain. One of the procedures employed with the framework of this second method (Cerven, E. et.al., Upsala J. Med. Sci. 86 xx (1981) p. 39-53) relies on the assumption that galactose is present as a terminal group of desialotransferrins. Cerven, E. et.al. therefore propose a procedure involving the use of a galactose-binding lectin for obtaining a biospecific affinity reaction between that lectin and the aforesaid assumed terminal galactose. At first a serum sample is contacted with an immunoadsorbent whereby transferrin--irrespective of its iso form--is caused to bind to said adsorbent. Then the transferrin-loaded adsorbent is separated, and the desialotransferrin attached thereto is to be detected with the aid of labeled galactose-binding lectin.
Both of these known methods involve heavy drawbacks. The IEF method is unpractical and takes too much time to be suitable for clinical routine work. In the course of our experiments for further developing a test according to Cerven, E. et.al. we have found that there is no correlation between the results obtained according to the lectin method and those obtained according to the IEF method. Obviously, therefore, these two methods do not measure the same items. It is not possible either, by means of t
REFERENCES:
patent: 4507390 (1985-03-01), Horiuchi et al.
patent: 4579661 (1986-04-01), Gustafsson et al.
Separation of Transferrin Types in Human Plasma by an Ion-Exchange High Performance Liquid Chromatography by Strahler et al. in Journal of Chromatography 266(1983), 281-291, Elsevier Scientific Pub. Amsterdam, Netherlands.
Quantification of a Transferrin Variant After Isoelectric Focusing in Agarose Gel Using Carbamylated Myoglobin as a Colored Marker by Petren et al. in Electrophoresis, vol. 5, pp. 26-29, Pub. 1984.
Quantative Analysis of Multiple Molecular Forms of Transferrin Using Isoelectric Focusing and Zone Immunoelectrophoresis Assay by Vesterberg et al. Journal of Immunological Methods, 46(1981), 53-62.
Chromafocusing: A Method for High Resolution Protein Purification by Richey et al. American Laboratory, pp. 100-110, Oct. 1981.
Evidence of Reduced Sialic Acid Content in Serum Transferrin in Male Alcoholics by Stibler et al. Alcoholic Clinical and Experimental Research, Karolinska Hospital, Stockholm, Sweden, pp. 545-549, Fall 1981.
Glycan Uniformity Within Molecular Variants of Transferrin With Distinct Affinity for Concanavalin A by Kerckaert et al. Biochemical and Biophysical Research Communications, vol. 105, No. 3, pp. 1023-1030, Apr. 1982, Academic Press, Lille Cedex, France.
Human Transferrin Asialotransferrin and the Intermediate Forms by Wong et al., McMasters University Health Science Center, Hamilton, Ontario, Canada, pp. 27-37, 1978.
Blanche Claes G.
Joustra Marius K.
Adee John
Pharmacia AB
Philpitt Fred
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