公开(公告)号：WO01049745A1

公开(公告)日：2001-07-12

申请号：PCT/US2000/035600

申请日：2000-12-29

IPC: G01N33/53 ,  B01J20/281 ,  C07K16/18 ,  C07K16/44 ,  C12M1/00 ,  C12N5/10 ,  C12N15/09 ,  C12P21/08 ,  C12Q1/68 ,  G01N30/88 ,  G01N37/00 ,  G01N33/531 ,  G01N33/552 ,  G01N33/577

CPC classification number: C07K16/44 ,  B01J2219/00274 ,  C07K16/18 ,  C12Q1/6804 ,  Y02P20/55

Abstract: This application describes an antibody that specifically binds to a synthetic oligomer (e.g., an oligonucleotide or oligopeptide) having a organic protecting group covalently bound thereto, which antibody does not bind to that synthetic oligomer when the organic protecting group is not covalently bound thereto. Methods of making and using such antibodies are also disclosed, along with cells for making such antibodies and articles carrying immobilized oligomers that can be used in assay procedures with such antibodies.

Abstract translation: 本发明涉及特异性结合的合成低聚物（如寡核苷酸或寡肽），其共价键合的基团的有机保护的抗体，所述抗体不结合该合成寡聚体时，有机保护基团 不与其共价结合。 本发明还涉及一种方法，用于获得和使用这样的抗体，以及对涉及生产这种抗体和物品的携带固定化低聚物可以用作的方法的一部分的细胞 用这种抗体进行分析。

公开(公告)号：WO01048480A1

公开(公告)日：2001-07-05

申请号：PCT/US2000/035583

申请日：2000-12-28

IPC: G01N33/53 ,  B01D15/08 ,  C07H21/02 ,  C07K1/22 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/566 ,  G01N33/68

CPC classification number: G01N33/5008 ,  C07H21/02 ,  C12Q1/6804 ,  C12Q1/6809 ,  C12Q2600/158 ,  G01N33/5023 ,  G01N33/5308 ,  G01N33/68 ,  G01N33/6803 ,  G01N33/6845 ,  C12Q2522/101

Abstract: Cellular mRNA-protein (mRNP) complexes are partitioned in vivo by contacting a biological sample with at least one ligand that specifically binds at least one component of a mRNP complex. Suitable biological samples comprise at least one mRNA-protein (mRNP) complex and include cell cultures, cell extracts, and whole tissue, including tumor tissue. Ligands include antibodies that specifically bind RNA-binding or RNA-associated proteins present in the mRNP complex. The mRNP complex is separated by binding the ligand with a binding molecule specific for the ligand, where the binding molecule is attached to a solid support. The mRNP complex is collected by removing the mRNP complex from the solid support. After collecting the mRNP complex, the mRNA bound within the complex may be characterized and identified. Subsets of the total mRNA population of a cell may accordingly be characterized, and a gene expression profile of the cell obtained.

Abstract translation: 通过使生物样品与至少一种特异性结合mRNP复合物的至少一种组分的配体接触，在体内分配细胞mRNA-蛋白（mRNP）复合物。 合适的生物样品包含至少一种mRNA-蛋白（mRNP）复合物，并且包括细胞培养物，细胞提取物和包括肿瘤组织在内的整个组织。 配体包括特异性结合存在于mRNP复合物中的RNA结合或RNA相关蛋白的抗体。 通过将配体与对配体特异性的结合分子结合来分离mRNP复合物，其中结合分子连接到固体支持物上。 通过从固体支持物上除去mRNP复合物来收集mRNP复合物。 在收集mRNP复合物后，可以鉴定和鉴定复合物内结合的mRNA。 因此可以对细胞的总mRNA群体的亚群进行表征，并获得所获得的细胞的基因表达谱。

公开(公告)号：WO01036681A2

公开(公告)日：2001-05-25

申请号：PCT/US2000/031277

申请日：2000-11-14

IPC: G01N31/22 ,  C07K16/44 ,  C12N15/09 ,  C12N15/11 ,  C12Q1/68 ,  G01N33/53 ,  G01N33/543 ,  G01N33/577 ,  G01N37/00

CPC classification number: C12Q1/6804 ,  C12Q1/6837 ,  C12Q2565/537 ,  C12Q2521/107 ,  C12Q2563/131

Abstract: The present invention provides a hybridization method and kit for the detection and measurement of biological molecules. A test sample containing the biological molecules of interest is hybridized with an unlabeled or optionally a detectably labeled complementary biomolecule to form a double-stranded hybrid immobilized to a solid phase. The immobilized hybrid may be detected with an entity which specifically recognizes an RNA:DNA hybrid, followed by analyses and quantification. Therefore, the present invention provides a method and kit to detect and measure biological molecules that is simple to use, highly specific, sensitive, and accurate for screening a plurality of biological molecules.

Abstract translation: 本发明提供了用于检测和测量生物分子的杂交方法和试剂盒。 将含有生物分子的测试样品与未标记或任选的可检测标记的互补生物分子杂交以形成固定于固相的双链杂交。 可以用特异性识别RNA：DNA杂交体的实体检测固定化的杂交，然后分析和定量。 因此，本发明提供了一种用于检测和测量生物分子的方法和试剂盒，所述生物分子易于使用，高度特异性，灵敏和精确地筛选多个生物分子。

公开(公告)号：WO00068434A2

公开(公告)日：2000-11-16

申请号：PCT/US2000/012391

申请日：2000-05-05

IPC: G01N33/53 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/532 ,  G01N33/566 ,  G01N33/58

CPC classification number: C12Q1/6804 ,  C12Q1/682 ,  C12Q2565/632 ,  C12Q2531/125

Abstract: Disclosed is a method of detecting multiple analytes in a sample in a single assay. The method is based on encoding target molecules with signals followed by decoding of the encoded signal. This encoding/decoding uncouples the detection of a target molecule from the chemical and physical properties of the target molecule. In basic form, the disclosed method involves association of one or more reporter molecules with one or more target samples, association of one or more decoding tags with the reporter molecules, and detection of the decoding tags. The reporter molecules associate with target molecules in the target sample(s). Generally, the reporter molecules correspond to one or more target molecules, and the decoding tags correspond to one or more reporter molecules. Thus, detection of particular decoding tags indicates the presence of the corresponding reporter molecules. In turn, the presence of particular reporter molecules indicates the presence of the corresponding target molecules. The sensitivity of the disclosed method can also be enhanced by including a signal amplification step prior to detection. Medical applications of this method include the analysis of the phenotypic status or replicative status of cells (growth or quiescence) and the assessment of normal and neoplastic cells in histologic or cytologic specimens in normal and disease states. For example, a pathologist may use the method to link a phenotypic state with the protein profile of lesion believed to contain malignant or pre-malignant cells.

Abstract translation: 公开了在单个测定中检测样品中的多种分析物的方法。 该方法基于编码具有信号的目标分子，随后对编码信号进行解码。 这种编码/解码将目标分子的检测与目标分子的化学和物理性质分离。 在基本形式中，所公开的方法包括将一个或多个报告分子与一个或多个目标样品相关联，一个或多个解码标签与报告分子的缔合以及解码标签的检测。 报告分子与目标样品中的靶分子缔合。 通常，报告分子对应于一个或多个目标分子，并且解码标签对应于一个或多个报告分子。 因此，特定解码标签的检测表明存在相应的报告分子。 反过来，特定报道分子的存在表明存在相应的靶分子。 所公开的方法的灵敏度还可以通过在检测之前包括信号放大步骤来增强。 该方法的医学应用包括分析正常和疾病状态下组织学或细胞学标本中细胞（生长或静止）的表型状态或复制状态以及正常和肿瘤细胞的评估。 例如，病理学家可以使用该方法将表型状态与相信含有恶性或恶性前细胞的病变的蛋白质谱相链接。

公开(公告)号：WO00060116A1

公开(公告)日：2000-10-12

申请号：PCT/US2000/008465

申请日：2000-03-31

IPC: C12Q1/68 ,  C40B40/06 ,  C40B60/14

CPC classification number: C12Q1/6804 ,  B01J2219/00317 ,  B01J2219/005 ,  B01J2219/00504 ,  B01J2219/00527 ,  B01J2219/00585 ,  B01J2219/00596 ,  B01J2219/00605 ,  B01J2219/0061 ,  B01J2219/00621 ,  B01J2219/00626 ,  B01J2219/0063 ,  B01J2219/00637 ,  B01J2219/00659 ,  B01J2219/00707 ,  B01J2219/00722 ,  C12Q1/6834 ,  C12Q1/6837 ,  C40B40/06 ,  C40B60/14 ,  C12Q2565/518 ,  C12Q2563/149

Abstract: The present invention relates to methods for performing high-throughput biological assays, including immunoassays, nucleic acid detection assays, and related assays. In a preferred embodiment, the target analyte to be detected is a specific mRNA. In another embodiment, the assay monitors the impact of a particular substance or effector on transcription. Alternatively, the target analyte may be a protein, hapten, non-biological chemical, pharmaceutical or other target material. The assay provides high amplification to maximize signal, and utilizes samples which do not require considerable purification.

Abstract translation: 本发明涉及用于进行高通量生物测定的方法，包括免疫测定，核酸检测测定和相关测定。 在优选的实施方案中，待检测的靶分析物是特异性mRNA。 在另一个实施方案中，测定监测特定物质或效应物对转录的影响。 或者，目标分析物可以是蛋白质，半抗原，非生物化学品，药物或其它靶材料。 该测定法提供高扩增以使信号最大化，并利用不需要相当多纯化的样品。

公开(公告)号：WO0018962A9

公开(公告)日：2000-08-24

申请号：PCT/US9922584

申请日：1999-09-28

Applicant: AFFYMETRIX INC ,  GOLDBERG MARTIN J ,  YELAGALAWADI GOVINDA RAO S ,  TANIMOTO EUGENE YUJI ,  TRAN HUU MINH ,  DONG HELIN ,  LOCKHART DAVID J ,  RYDER THOMAS B ,  WARRINGTON JANET A ,  BEECHER JODY

Inventor： GOLDBERG MARTIN J ,  YELAGALAWADI GOVINDA RAO S ,  TANIMOTO EUGENE YUJI ,  TRAN HUU MINH ,  DONG HELIN ,  LOCKHART DAVID J ,  RYDER THOMAS B ,  WARRINGTON JANET A ,  BEECHER JODY

IPC: C12N15/09 ,  C12Q1/68 ,  G01N37/00 ,  G01N33/58

CPC classification number: C12Q1/682 ,  C12Q1/6804 ,  Y10S436/807 ,  C12Q2565/501 ,  C12Q2563/131 ,  C12Q2527/125 ,  C12Q2563/107 ,  C12Q2563/125 ,  C12Q2563/143 ,  C12Q2537/125

Abstract: Methods and compounds are provided for detecting target molecules in a sample using specific binding assays. In particular, methods are provided for detecting a nucleic acid target in a sample. In one embodiment, the method comprises hybridizing a nucleic acid target, comprising a target nucleic acid sequence, to a nucleic acid probe, comprising a probe nucleic acid sequence, wherein the target comprises a binding ligand. The hybridized target is contacted with a receptor comprising multiple sites capable of binding the binding ligand to complex the receptor to the binding ligand, and the receptor is contacted with an amplification reagent, comprising a plurality of the binding ligands, to complex the amplification reagent to the receptor. The presence of the complexed amplification reagent then is detected, for example, by detecting the presence of a detectable label, such as a fluorescent label, for example, on the receptor or the amplification reagent. Optionally, the amplification reagent, comprising a plurality of the binding ligands, is contacted with labeled receptor molecules thereby to complex a plurality of labeled receptor molecules to the amplification reagent, and the labeled receptor molecules complexed to the amplification reagent are detected. This permits the detectable signal to be enhanced and amplified. In one embodiment, the binding ligand is biotin, the receptor is streptavidin, and the amplification reagent is an antibody or a DNA matrix. In another embodiment, an array of different nucleic acid probes immobilized on a surface, each having a defined sequence and location on the surface, may be used in the assays, thus permitting screening and detection of binding of large number of nucleic acids.

Abstract translation: 提供了使用特异性结合测定法检测样品中的靶分子的方法和化合物。 具体而言，提供了用于检测样品中的核酸靶标的方法。 在一个实施方案中，所述方法包括使包含靶核酸序列的核酸靶与包含探针核酸序列的核酸探针杂交，其中所述靶包含结合配体。 使杂交的靶与包含能够结合结合配体的多个位点的受体接触以将受体与结合配体复合，并使受体与包含多个结合配体的扩增试剂接触以将扩增试剂复合成 受体。 然后检测复合的扩增试剂的存在，例如通过检测例如受体或扩增试剂上的可检测标记（例如荧光标记）的存在。 任选地，使包含多个结合配体的扩增试剂与标记的受体分子接触，从而将多个标记的受体分子与扩增试剂复合，并检测与扩增试剂复合的标记的受体分子。 这允许可检测信号被增强和放大。 在一个实施方案中，结合配体是生物素，受体是链霉抗生物素蛋白，扩增试剂是抗体或DNA基质。 在另一个实施方案中，固定在表面上的不同核酸探针的阵列，每个核酸探针在表面上都具有限定的序列和位置，可以用于测定中，从而允许筛选和检测大量核酸的结合。

公开(公告)号：WO00022165A1

公开(公告)日：2000-04-20

申请号：PCT/GB1999/003383

申请日：1999-10-12

IPC: G01N33/53 ,  C12N15/09 ,  C12Q1/34 ,  C12Q1/68 ,  G01N33/538 ,  G01N33/566 ,  G01N33/58 ,  G01N37/00

CPC classification number: C12Q1/6832 ,  C12Q1/6804 ,  C12Q2563/125 ,  C12Q2531/113 ,  C12Q2527/119 ,  C12Q2565/531 ,  C12Q2525/107 ,  C12Q2521/307 ,  C12Q2522/101

Abstract: The present invention provides a method for detecting a single-stranded target nucleic acid comprising the steps of: a) forming a hybrid between a target nucleic acid and a nucleic acid probe, said nucleic acid probe labelled with an enzyme reagent which hydrolyses single-stranded nucleic acid but is substantially without effect on double-stranded nucleic acid, said hybrid formed under conditions of pH which are outside the activity range of said enzyme reagent; b) adjusting said pH to a value within the activity range of said enzyme reagent, whereby said enzyme reagent substantially hydrolyses any single-stranded nucleic acid present; and c) contacting said hybrid with a detection reagent to detect the hybrid, characterised by , prior to step (c), bringing the nucleic acid probe or hybrid into contact with a solid support to attach it thereto or bringing the nucleic acid probe or hybrid into contact with a capture reagent, optionally linked to a solid support, to capture the nucleic acid probe or hybrid; and washing the capture reagent or solid support on which the hybrid is immobilised with a washing fluid while the capture reagent or solid support is contained within a vessel that is adapted to retain the capture reagent or solid support but not to retain fluid in which the capture reagent or solid support is dispersed, whereby material which has not been captured by the capture reagent or otherwise immobilised on a solid support is eluted from the vessel.

Abstract translation: 本发明提供了检测单链靶核酸的方法，包括以下步骤：a）在靶核酸和核酸探针之间形成杂交体，所述核酸探针用酶水解标记的核酸探针，其水解单链 核酸，但对双链核酸基本上没有影响，所述杂交体在pH范围内在不在所述酶试剂的活性范围之外的条件下形成; b）将所述pH调节至所述酶试剂的活性范围内的值，由此所述酶试剂基本上水解存在的任何单链核酸; 和c）在步骤（c）之前使所述杂交体与检测试剂接触以检测特征在于的杂交体，使核酸探针或杂交体与固体支持物接触以将其附着于其上，或 使核酸探针或杂交体与捕获试剂（任选地连接到固体支持物）接触以捕获核酸探针或杂交体; 并且在捕获试剂或固体支持物被包含在适于保持捕获试剂或固体支持物但不保留其中捕获的流体的容器内时，用洗涤液洗涤固定有杂交体的捕获试剂或固体载体 分散试剂或固体支持物，由此未被捕获试剂捕获或以其他方式固定在固体载体上的物质从容器中洗脱出来。

公开(公告)号：WO9958971A3

公开(公告)日：2000-01-20

申请号：PCT/CA9900444

申请日：1999-05-13

Applicant: LE XIAO CHUN ,  WEINFELD MICHAEL ,  XING JAMES Z

Inventor： LE XIAO-CHUN ,  WEINFELD MICHAEL ,  XING JAMES Z

IPC: C12Q1/68 ,  G01N33/48

CPC classification number: C12Q1/6804 ,  C12Q2565/137

Abstract: The present invention provides methods for the detection and quantitation of any modification of interest in any nucleic acid sequence. In particular, the invention provides methods for detecting and quantitating low levels of modifications of interest in DNA sequences. The methods of the invention take advantage of combining the use of nucleic acid sequence modification-specific molecules which are specific for DNA modifications, of fluorescently labelled proteins which are specific for the nucleic acid sequence modification-specific molecules, of capillary electrophoresis and of laser-induced fluorescence. The methods of the invention are useful for identifying and detecting exposure to carcinogens, in early risk assessment for cancer, and in monitoring of cancer therapy.

Abstract translation: 本发明提供了用于检测和定量任何核酸序列的兴趣修饰的方法。 特别地，本发明提供了检测和定量DNA序列中低水平的感兴趣修饰的方法。 本发明的方法利用了将对DNA修饰特异的核酸序列修饰特异性分子，对核酸序列修饰特异性分子特异性的荧光标记蛋白，毛细管电泳和激光 - 诱导荧光。 本发明的方法可用于鉴定和检测致癌物的暴露，癌症的早期风险评估和癌症治疗的监测。

公开(公告)号：WO99029892A1

公开(公告)日：1999-06-17

申请号：PCT/US1998/025767

申请日：1998-12-04

IPC: C12Q1/04 ,  C12Q1/66 ,  C12Q1/68 ,  C12Q1/70 ,  G01N33/53

CPC classification number: C12Q1/689 ,  C12Q1/66 ,  C12Q1/6804 ,  C12Q1/6806 ,  C12Q1/6895 ,  C12Q1/70 ,  G01N33/5308 ,  Y02A50/52

Abstract: The presence or absence of an organism in a sample is detected by isolating the organism from the sample by a suitable affinity matrix, releasing the organism from the affinity matrix, rupturing the cells of the organism to release total nucleic acid and hydrolyzing or digesting the total nucleic acid to form mononucleotides or individual free nucleic acid bases and inorganic phosphate to form an analyte solution, and assaying the analyte solution for the at least one presence of free nucleic acid base or inorganic phosphate to thereby determine whether the organism was present in the sample.

Abstract translation: 通过用合适的亲和基质从样品中分离生物体来检测样品中有机体的存在或不存在，从亲和基质释放生物体，破坏生物体的细胞以释放总核酸并水解或消化总量 核酸以形成单核苷酸或单个游离核酸碱基和无机磷酸盐以形成分析物溶液，以及测定分析物溶液至少存在游离核酸碱基或无机磷酸盐，从而确定生物体是否存在于样品中 。

公开(公告)号：WO99004042A1

公开(公告)日：1999-01-28

申请号：PCT/US1998/014782

申请日：1998-07-16

IPC: C12Q1/68 ,  G01N33/543 ,  G01N33/569 ,  G01N33/58 ,  C07H21/04 ,  G01N33/53

CPC classification number: G01N33/58 ,  C12Q1/6804 ,  C12Q1/6834 ,  G01N33/54326 ,  G01N33/569 ,  C12Q2565/531

Abstract: Compositions and methods for detecting or purifying multiple selected nucleic acid molecules are provided. The compositions comprise oligonucleotide-peptide conjugates (OPCs) and antibodies immunologically specific for the conjugates. The oligonucleotide moiety of each conjugate specifically hybridizes with a selected target nucleic acid molecule. The peptide moiety acts as an antigen for generation of a specific antibody that uniquely binds to its respective OPC. OPCs are advantageously used in groups, for detection or separation of multiple target nucleic acids in a single test sample, by virtue of the unique antigen/antibody interaction that enables differential detection or capture of the OPC. Methods and kits are provided for using the OPC systems of the invention.

Abstract translation: 提供了用于检测或纯化多个选择的核酸分子的组合物和方法。 组合物包含寡核苷酸 - 肽缀合物（OPCs）和针对缀合物免疫特异性的抗体。 每个缀合物的寡核苷酸部分与选定的靶核酸分子特异性杂交。 肽部分用作用于产生唯一结合其相应OPC的特异性抗体的抗原。 通过独特的抗原/抗体相互作用，OPCs有利地用于组中，用于在单个测试样品中检测或分离多个靶核酸，其能够进行OPC的差异检测或捕获。 提供了使用本发明的OPC系统的方法和试剂盒。