1/301335 - (C) Derwent AN - 93-182698 [22] XA - C93-080980 XP - N93-140416 TI - Quantitating presence or activity of enzyme - by incubating with modified peptide

substrate and measuring the modified peptide prod. PA - (PROM-) PROMEGA CORP IN - SHULTZ J W; WHITE D H DC - B04 D16 S03 NP - 1 NC - 036 PN - WO9310461 A1 930527 DW9322 G01N-033/573 Eng 103pp DS - AT BE CH DE DK ES FR GB GR IE IT LU MC NL OA SE DN - AT AU BB BG BR CA CH CS DE DK ES FI GB HU JP KP KR LK LU MG MN MW NL NO PL RO RU SD SE PR - 91US-791928 911112 AP - 92WO-U09595 921112 CT - 3.Jnl.Ref; J02240027; US4681842; US4687732; US4803157; US4912033 ; WO8500809 IC - C12Q-001/42; G01N-033/53; G01N-033/573 AB - WO9310461 Method comprises (a) adding a modified peptide substrate to a soln. contg. the enzyme, (b) incubating the enzyme with the modified peptide substrate under conditions where the enzyme is active for a time sufficient to form a modified peptide prod., (c) sepg. the modified peptide prod. from the modified peptide substrate and (d) measuring the quantity of modified peptide prod. The modified peptide substrate may include a detector segment, e.g. dansyl, sulphorhodamine, lissamine rhodamine (rhodamine B) or fluorescein. USE/ADVANTAGE - The methods and reagents can be used for the rapid, specific and sensitive detection of enzymes such as protein kinases, phosphatases and proteases. They can be used to study enzyme function in metabolism and in diagnosis of disease. (Dwg. 0/20) 2/301335 - (C) Derwent AN - 93-182697 [22] XA - C93-080979 XP - N93-140415 TI - Rapid, highly sensitive assay of polychlorinated dioxin - by detecting protein tyrosine phosphorylation, useful for phenotypic tumour cells partic. breast and colon cancer cells PA - (CORR ) CORNELL RES FOUND INC IN - BABISH J G; EL-TOM S; MA X; WHEELOCK G DC - B02 B04 S03 NP - 1 NC - 016 PN - WO9310460 A1 930527 DW9322 G01N-033/567 Eng 048pp DS - AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE DN - JP PR - 91US-792922 911115; 91US-795193 911119 AP - 92WO-U09765 921113 CT - 7.Jnl.Ref IC - G01N-033/567 AB - WO9310460 To detect, in a sample, the presence of polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), and their structural analogues, which exhibit biological activity characteristic of PCDDs comprising prodn. of phosphotyrosyl proteins (PTPs) in hepatocytes of female C57BL/6 mice, or other strain with cytochrome P-450IA1 inducibility, comprises (a) admin. of the test sample to the mice; and (b) examining the hepatic S-9 supernatant for

phosphorylation of tyrosine residues of S-9 proteins. Also new is an in vitro bioassay using the above method; but using X3 lymphoblastoma cells or other cell line with cytochrome P-450IA1 inducibility, comprising admin. of the test sample to the cell culture and examining the cell lysate for phosphorylation of tyrosine residues of certain proteins. USE/ADVANTAGE - Method provides a low-cost, rapid immunoassay sensitive to parts per trillion or billion, with high selectivity and without necessity for a preliminary clean-up to remove interfering substances. A further use is for phenotyping of tumour cells from a biopsy sample, partic. breast and colon cancer cells, by treatment with low concns. of TCDD and assessing the increase in protein tyrosine phosphorylation (pTP), as mediated by the aryl hydrocarbon (Ah) receptor. (Dwg.11/13)