CELL BASED QUALITY CONTROL BIOASSAYS FOR NUTRICEUTICAL AND MEDICINAL PRODUCTS Russian patent published in 2018 - IPC G01N33/50 

Abstract RU 2662358 C2

FIELD: measurement; testing.

SUBSTANCE: method comprises: contacting a line cell that has an access number of PTA-13010 in the American Type Culture Collection (ATCC) with said composition for a period of time and at a temperature sufficient to inhibit the proliferation of said cell, contacting a cell line that has an access number of PTA-13011 in the ATCC with said composition for a period of time and at a temperature sufficient to inhibit the proliferation of said cell, measuring the proliferation inhibition level of the said cell line induced by said composition, which has an access number of PTA-13010 in the ATCC, and a proliferation inhibition level of said cell line which has an access number of PTA-13011 in the ATCC, wherein the amount of said inhibitory activity of said composition with respect to the translation initiation process is proportional to the proliferation inhibition level in the cell of the line that has an access number of PTA-13010 in the ATCC, and comparing the proliferation inhibition level induced by said composition with the proliferation inhibition level induced by a standard having a known amount of said activity, and determining the composition as having no inhibitory activity with respect to translation initiation if said composition inhibits the proliferation of said cell, a line that has an access number of PTA-13011 in the ATCC; wherein said composition is a food, nutraceutical or medicinal product, wherein said cell of the line that has an access number of PTA-13010 and / or PTA-13011 in the ATCC is characterised in that it contains a lentiviral expression vector that contains: (a) a heterologous eIF2α gene, which is completely free of 3' and 5' NTO; (b) directed at nucleotide No. 1098, thymidine nucleotide in 3'-NTO areas of the endogenous eIF2 gene, siRNA No. 1098, which inhibits more than 85% of the endogenous expression of eIF2; and (c) Kozak optimal consensus sequence (GCCACCATGG). Invention also relates to: human prostate cancer cell lines PC-3, expressing eIF2α-WT, which has an access number of PTA-13010 in the ATCC, in which the expression of endogenous eIF2α is inhibited by more than 85%, and human prostate cancer cell lines PC-3, expressing eIF2α-S51A, which has an access number of PTA-13011 in the ATCC, in which the expression of endogenous eIF2α is inhibited by more than 85%, to determine inhibition of translation initiation.

EFFECT: invention relates to a method for determining the effectiveness of a composition with respect to inhibition of translation initiation, the level of inhibitory activity of which with respect to translation initiation is unknown.

19 cl, 4 dwg, 6 ex

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RU 2 662 358 C2

Authors

Khalperin Zhoze A.

Chorev Majkl

Aktas Khusejn

Dates

2018-07-25Published

2013-07-22Filed