Test for oxidative stress using cell suspensions

Details Test for oxidative stress using cell suspensions Test for oxidative stress using cell suspensions

2005-02-22

2005-02-22

Chin, Christopher L. (Department: 1641)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

C435S002000, C435S004000, C435S007250, C435S025000, C435S375000, C435S383000, C435S384000, C436S503000, C436S063000, C436S071000, C436S901000

Reexamination Certificate

active

06858399

ABSTRACT:
This invention provides a method of measuring oxidative response of cells without recourse to preparation of cell culture. The process involves:1) preparing suspensions of cells from a living host in isotonic solutions,2) preparing samples of test materials in isotonic solution containing tagged choline,3) adding the cells suspension prepared in step 1 to the samples prepared in step 2,4) incubating the product of step 3 with shaking for 2-90 minutes,5) extracting and drying the lipid phase from the product of step 4, and6) subjecting the product of step 5 to a scintillation counter to measure choline which has been incorporated into phosphatidylcholine (PC).An increase in incorporation of choline into PC in the short term indicates oxidative stress or free radical induced damage. Because the method of the invention using the cell isolates does not require the expense of cell culture with concomitant expense and possibility of cell change, it is particularly useful for clinical evaluation. Furthermore, the ability to use erythrocytes for study of cellular response to oxidative stress makes it possible to test individual responses of patient using part of the blood sample drawn routinely for laboratory use.

REFERENCES:
patent: 6218130 (2001-04-01), Lamb
Wertz et al., 1980. Inhibition of 12-O-tetradecanoylphorbol-13-acetate-accelerated phospholipid metabolism by 5,8,11,14-eicosatetraynoic acid. Cancer Res. 40: 776-781.*
Anderson et al., 1985. Role of fatty acid structure in the reversible activation of phosphatidylcholine synthesis in lymphocytes. Biochimica et Biophysica Acta 835: 360-368.*
Pacifici et al., 1994. Lipid hydroperoxide-induced peroxidation and turnover of endothelial cell phospholipids. Free Radical Biol. Med. 17: 297-309.*
Ferrali et al., 1989. Allyl alcohol-induced hemolysis and its relation to iron release and lipid peroxidation. Biochem. Pharmacol. 38: 1819-1825.*
Strunecka et al., 1988. Phospholipid biosynthesis in mature human erythrocytes. In, Gen. Physiol. Biophys. 7: 205-216. Abstract, BIOSIS No. 86000720.

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