公开(公告)号：US06962808B2

公开(公告)日：2005-11-08

申请号：US09910628

申请日：2001-07-20

申请人： Judith L. Erb ,  James G. Downward, IV ,  John R. Erb-Downward ,  James L. Witliff

发明人： Judith L. Erb ,  James G. Downward, IV ,  John R. Erb-Downward ,  James L. Witliff

IPC分类号： G01N21/77 ,  C12M1/34

CPC分类号： G01N21/648 ,  G01N21/7703 ,  Y10S435/808 ,  Y10S436/805 ,  Y10S436/811 ,  Y10S436/813 ,  Y10S436/814 ,  Y10S436/815 ,  Y10S436/817

摘要： There is provided an apparatus for screening pharmacological agents for agents which induce regression of cancer. The apparatus includes an evanescent sensing device, at least one sensor having affixed to its surface molecules of a first type, which have affinity for molecules of a biological receptor, the surface molecule and receptor molecule combination having the effect that, in vivo, the binding affects the rate of transcription of gene products, and a molecular tag wherein the molecular tag is bound to the sensor wherein the binding between molecules of the first type and molecules the biological receptor cause the tag to produce a alteration in signal recorded by the evanescent sensing device, the tag also being bound to molecules of a second type, the molecules of the second type having affinity for the receptor molecules. Also provided is a method for screening pharmacological agents to determine agents which induce regression of cancer by contacting extract from a tumor tissue biopsy with a molecular tag, thereby causing the tag to bind to receptor molecules present in the tumor tissue biopsy, flowing the tag sample extract through a sensor, as set forth above, and recording the time course of signal observed by the evanescent sensing device, introducing pharmacological agents to be accessed to the tag sample extract, flowing the tag sample extract through the sensor again and recoding the time course of signal observed by the evanescent sensing device, and using the data to evaluate the impact of the pharmacological agents on the rate of transcription of gene products.

摘要翻译： 提供了用于筛选诱导癌症消退的药剂的药理学试剂的装置。 所述装置包括消逝传感装置，至少一个传感器固定在其表面上具有第一类型的分子，其具有对生物受体分子的亲和力，所述表面分子和受体分子组合具有在体内结合 影响基因产物的转录速率和分子标签，其中分子标签与传感器结合，其中第一类型的分子与分子之间的结合生物受体导致标签产生由ev逝感测记录的信号的变化 该标签也与第二类型的分子结合，第二类型的分子对受体分子具有亲和力。 还提供了一种用于筛选药理学试剂以确定通过将来自肿瘤组织活检的提取物与分子标签接触来诱导癌症消退的试剂的方法，从而使标签与存在于肿瘤组织活组织检查中的受体分子结合，使标签样品 通过如上所述的传感器提取并记录由ev逝感测装置观察到的信号的时间过程，将待接入的药剂引入标签样品提取物，再次使标签样品提取物再次通过传感器并重新编码时间过程 的消逝感测装置观察到的信号，并使用数据来评估药理学试剂对基因产物转录速率的影响。

公开(公告)号：US06300082B1

公开(公告)日：2001-10-09

申请号：US09045224

申请日：1998-03-20

申请人： Judith L. Erb ,  James G. Downward, IV ,  John R. Erb-Downward ,  James L. Wittliff

发明人： Judith L. Erb ,  James G. Downward, IV ,  John R. Erb-Downward ,  James L. Wittliff

IPC分类号： G01N33543

CPC分类号： G01N21/648 ,  G01N21/7703 ,  Y10S435/808 ,  Y10S436/805 ,  Y10S436/811 ,  Y10S436/813 ,  Y10S436/814 ,  Y10S436/815 ,  Y10S436/817

摘要： A method and apparatus for measuring binding between a plurality of molecules of a biological receptor protein and a plurality of molecules of a type which binds to said biological receptor is presented. Apparatus utilizes a sensor possessing a waveguide to which have been attached in close proximity to its surface, features resembling molecules having binding affinity for said biological receptor. Light is injected into said waveguide so as to produce an evanescent field at its surface. Molecules of receptor protein 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 the waveguide as binding proceeds between tagged receptor protein and the binding molecular feature of the waveguide. This allows direct measurement of binding and dissociation rates between the receptor and the binding feature of the waveguide. By using a waveguide having a feature resembling a ligand for the receptor, the potential hormonal activity of a test substance may be evaluated from its ability to compete with the waveguide for binding with the receptor. The effect of a test compound on binding of receptor protein to a subsequent element in a hormonal signal transduction mechanism is evaluated by measuring the impact of the test compound on binding between receptor protein and a feature resembling said next element of the signal transduction mechanism. 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 receptor and test ligand and for the receptor and the next element of the signal transduction mechanism. 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 estrogen mimics present in a test sample, and for evaluation of pharmaceuticals intended to treat hormone dependent cancers.

公开(公告)号：US06251688B1

公开(公告)日：2001-06-26

申请号：US09045223

申请日：1998-03-20

申请人： Judith L. Erb ,  James G. Downward ,  John R. Erb-Downward ,  Otho Ulrich

发明人： Judith L. Erb ,  James G. Downward ,  John R. Erb-Downward ,  Otho Ulrich

IPC分类号： G01N33543

CPC分类号： G01N21/648 ,  G01N21/552 ,  G01N21/6428 ,  G01N21/7703 ,  G01N2021/6484 ,  G01N2021/7786 ,  Y10S435/808 ,  Y10S436/805 ,  Y10S436/814 ,  Y10S436/817 ,  Y10S436/818 ,  Y10S977/924 ,  Y10S977/958 ,  Y10S977/959

摘要： 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.

摘要翻译： 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，以便在其表面产生渐逝场。 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的化学品的标签标记，所述标签改变光的特性。 装置利用快速的方式监测来自所述波导的光信号的变化，因为在所述波导上标记的类型2的分子和类似于类型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。 提供了这样的方法，其中这样的数据可以用于计算亲和常数，结合活性，由协同性产生的复合亲和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应答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，以及用于评估组织活检样品以检测p53蛋白的突变体形式。