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    • 2. 发明公开
    • Method and apparatus for measuring binding between biological molecules
    • 用于测量生物分子之间的结合的方法和装置
    • EP0957354A3
    • 2000-03-22
    • EP99302175.7
    • 1999-03-22
    • IA Inc.
    • Erb,Judith L.Downward,James G.Erb-Downward,John R.Ulrich,OthoWitliff,James G.
    • G01N21/77G01N33/543
    • G01N21/648G01N21/7703
    • A method and apparatus for measuring binding between a plurality of molecules of a first type and a plurality of molecules of a second type is presented. Apparatus utilizes a sensor possessing a waveguide to which have been attached in close proximity to its surface, features resembling molecules of said first type. Light is injected into said waveguide so as to produce an evanescent field at its surface. Molecules of said second type are tagged with a tag belonging to that class of chemicals which alters a characteristic of light, when said light passes through said chemical tag. Apparatus utilizes a rapid means of monitoring the change in optical signal coming from said waveguide as binding proceeds between tagged molecules of type 2 and the feature resembling molecules of type 1 on said waveguide. This allows direct measurement of binding and dissociation rates between the two types of molecules. Methods are provided whereby such data may be used to compute affinity constants, binding activity, complex affinity constants resulting from cooperativity, and kinetic parameters for the molecular pair. Preferred embodiments of the invention illustrate application of the method and apparatus to measuring binding between biological receptors and their nuclear response elements, and the use of this type of measurement for assessment of the activity of hormonal mimics present in a sample, for evaluation of pharmaceuticals intended to treat hormone dependent cancers, and for evaluation of tissue biopsy samples to detect mutant forms of the p53 protein.
    • 5. 发明公开
    • Method and apparatus for measuring binding between biological molecules
    • Verfahren und Vorrichtung zum Messen der Bindung zwischenBiomolekülen
    • EP0957354A2
    • 1999-11-17
    • EP99302175.7
    • 1999-03-22
    • IA Inc.
    • The designation of the inventor has not yet been filed
    • G01N21/77
    • G01N21/648G01N21/7703
    • A method and apparatus for measuring binding between a plurality of molecules of a first type and a plurality of molecules of a second type is presented. Apparatus utilizes a sensor possessing a waveguide to which have been attached in close proximity to its surface, features resembling molecules of said first type. Light is injected into said waveguide so as to produce an evanescent field at its surface. Molecules of said second type are tagged with a tag belonging to that class of chemicals which alters a characteristic of light, when said light passes through said chemical tag. Apparatus utilizes a rapid means of monitoring the change in optical signal coming from said waveguide as binding proceeds between tagged molecules of type 2 and the feature resembling molecules of type 1 on said waveguide. This allows direct measurement of binding and dissociation rates between the two types of molecules. Methods are provided whereby such data may be used to compute affinity constants, binding activity, complex affinity constants resulting from cooperativity, and kinetic parameters for the molecular pair. Preferred embodiments of the invention illustrate application of the method and apparatus to measuring binding between biological receptors and their nuclear response elements, and the use of this type of measurement for assessment of the activity of hormonal mimics present in a sample, for evaluation of pharmaceuticals intended to treat hormone dependent cancers, and for evaluation of tissue biopsy samples to detect mutant forms of the p53 protein.
    • 提出了用于测量第一类型的多个分子与第二类型的多个分子之间的结合的方法和装置。 装置利用具有波导的传感器,其已经附着在其表面附近,特征类似于所述第一类型的分子。 光注入到所述波导中,以便在其表面产生消逝场。 当所述光通过所述化学标签时,所述第二类型的分子用属于所述类别化学品的标签标记,所述标签改变光的特性。 装置利用快速的方式来监测来自所述波导的光信号的变化,因为在所述波导上的类型2的标记分子和类似类型1的分子的特征之间的结合进行。 这允许直接测量两种类型的分子之间的结合和解离速率。 提供了方法,其中这样的数据可用于计算亲和常数,结合活性,由协同性产生的复合亲和力常数和分子对的动力学参数。 本发明的优选实施方案说明了用于测量生物受体及其核反应元件之间的结合的方法和装置的应用,以及使用这种类型的测量来评估样品中存在的激素模拟物的活性,以评估药物的目的 治疗激素依赖性癌症,以及用于评价组织活检样品以检测p53蛋白的突变体形式。