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1. 发明公开

KR1020070099519A 3가 금속 매개 균질 발광 근접 분석 无效
标题翻译：多金属介质均质光致亲和性测定
公开(公告)号：KR1020070099519A
公开(公告)日：2007-10-09
申请号：KR1020077007650
申请日：2005-09-19
申请人： 노바티스 백신즈 앤드 다이아그노스틱스 인코포레이티드
发明人： 쿠오리차드씨. , 더즈티모티디. , 마카쥬스키티모티디.
IPC分类号： C12Q1/48 , C12Q1/00 , G01N33/53
CPC分类号： G01N33/542 , C12Q1/42 , C12Q1/44 , C12Q1/485
摘要： An in vitro protein kinase assay technology that (1) exhibits a high assay signal to background ratio (S/B) and range (S-B); (2) is homogenous; (3) is non-radioactive; and (4) does not require a phospho-specific antibody involves complexing a trivalent metal ion (e.g. Ga3+, Fe3+, Al3+, In3+, Ru3+, Sc3+, Y3+) to the surface of amplified luminescent proximity assay acceptor or donor beads, e.g., via a suitable linker such as nitrilotriacetic acid (NTA; also referred to as carboxymethyl-lysine), iminodiacetic acid (IDA), or an appropriately substituted N-containing heterocycle, for example a triazoheterocycle, for example a triazocyclononaneononane, such as 1-propylamino-4-acetato-1,4,7-triazacyclononane. A protein (or constituent part) or other kinase substrate is bound to the surface of the other of an amplified luminescent proximity assay acceptor or donor bead and, if phosphorylated, brought into proximity with the trivalent metal ion-complexed acceptor bead to generate a luminescent signal. Presence of a kinase inhibitor inhibits phosphorylation and therefore signal generation and, in this way, is detectable. As the invention described herein recognizes the presence or absence of phosphate groups on a protein, (or constituent part), or other biological macromolecule (e.g., mono, di, or trinucleotides, cyclic nucleotides or phosphate substituted inositols), it is broadly applicable to any phosphorlylation or dephosphorylation reaction enzymes and provides a highly robust and flexible assay format for protein kinases and other enzyme classes, including lipid kinases, phosphatases, phosphodiesterases and others.
摘要翻译：（1）表现出高测定信号与背景比（S / B）和范围（S-B）的体外蛋白激酶测定技术; （2）是均匀的; （3）是非放射性的; 和（4）不需要磷酸特异性抗体涉及将三价金属离子（例如，Ga 3+，Fe 3+，Al 3+，In 3+，Ru 3+，Sc 3+，Y 3+）与扩增的发光亲和力测定受体或供体珠的表面络合，适当的接头如次氮基三乙酸（NTA;也称羧甲基赖氨酸），亚氨基二乙酸（IDA）或适当取代的含N的杂环，例如三氮杂环壬烷，例如三氮杂环壬烷，例如1-丙基氨基 - 4-乙酸基-1,4,7-三氮杂环。蛋白质（或组成部分）或其他激酶底物与扩增的发光亲和测定受体或供体小珠的另一个的表面结合，并且如果磷酸化，则与三价金属离子络合的受体珠接近以产生发光信号。激酶抑制剂的存在抑制磷酸化并因此产生信号，并且以这种方式可检测。如本文所述的发明认识到蛋白质（或组成部分）或其他生物大分子（例如单，二或三核苷酸，环状核苷酸或磷酸酯取代的肌醇）上磷酸基团的存在或不存在，广泛适用于任何磷酸化或去磷酸化反应酶，并为蛋白激酶和其他酶类提供了强大而灵活的测定形式，包括脂质激酶，磷酸酶，磷酸二酯酶等。

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