公开(公告)号：US5972721A

公开(公告)日：1999-10-26

申请号：US816429

申请日：1997-03-14

申请人： John G. Bruno ,  Johnathan L. Kiel ,  John P. Kilian

发明人： John G. Bruno ,  Johnathan L. Kiel ,  John P. Kilian

IPC分类号： B03C1/01 ,  B03C1/032 ,  B03C1/033 ,  B03C1/034 ,  G01N33/533 ,  B03C1/00

CPC分类号： B03C1/032 ,  B03C1/01 ,  B03C1/0332 ,  B03C1/034 ,  Y10S436/824

摘要： An apparatus and method for immunomagnetic separation and concentration of target biological materials is disclosed. The immunomagnetic separation is performed by a magnetic flow cell, or filter block, as part of an automated mostly continuous immunomagnetic assay system. The magnetic flow cell has two bundles of ferromagnetic rods or pins positioned inside an internal chamber so that a fluid sample flowing through the flow cell passes through the pins. A pair of cobalt magnets flank the flow cell so that the pins concentrate and sufficiently increase the magnetic fields so that even nanometer size magnetic beads can be captured. The overall system combines a reaction subsystem for reacting coated magnetic beads with a sample, a collection subsystem for capturing magnetic beads, a rinsing subsystem for removing debris and a filtering subsystem for removing captured magnetic beads from the collection subsystem. The new magnetic flow filter is the key component for the collection and filtering subsystems.

摘要翻译： 公开了一种用于免疫磁性分离和靶向生物材料浓缩的装置和方法。 免疫磁性分离由磁流量池或过滤块进行，作为自动化大多数连续的免疫磁性测定系统的一部分。 磁流量计具有两束位于内部室内的铁磁棒或销，使​​得流过流动池的流体样品通过销。 在流动池侧面的一对钴磁体，使得引脚集中并充分增加磁场，使得可以捕获甚至纳米尺寸的磁珠。 整个系统结合了用于使涂覆的磁珠与样品反应的反应子系统，用于捕获磁珠的收集子系统，用于去除碎屑的冲洗子系统和用于从收集子系统中去除捕获的磁珠的过滤子系统。 新的磁流过滤器是收集和过滤子系统的关键组件。

公开(公告)号：US06303316B1

公开(公告)日：2001-10-16

申请号：US09608706

申请日：2000-06-30

申请人： Johnathan L. Kiel ,  John G. Bruno ,  Jill E. Parker ,  John L. Alls ,  Charles R. Batishko ,  Eric A. Holwitt

发明人： Johnathan L. Kiel ,  John G. Bruno ,  Jill E. Parker ,  John L. Alls ,  Charles R. Batishko ,  Eric A. Holwitt

IPC分类号： C12Q168

CPC分类号： C12Q1/6825 ,  C12Q1/6837 ,  Y10T436/143333 ,  C12Q2565/607

摘要： In a recognition complex system, nucleic acid ligands comprising random DNA sequences are operatively coupled to an organic semiconductor and distributed so as to form an array of recognition complexes. When an unknown chemical or biological analyte is applied to the array, the electrical and/or photochemical properties of one or more of the recognition complexes are altered upon binding of the nucleic acid ligand to the analyte. The degree to which the electrical and/or photochemical properties change is a function of the affinity of the nucleic acid ligand sequence for the analyte. The electrical and photochemical changes associated with the array, as a whole, can be used as a unique signature to identify the analyte. In certain embodiments, an iterative process of selection and amplification of nucleic acid ligands that bind to the analyte can be used to generate a new array with greater affinity and specificity for a target analyte, or to produce one or more nucleic acid ligands with high binding affinity for an analyte. The present invention also provides methods for preparing nucleic acid ligands that bind with high affinity to an analyte and using such nucleic acid ligands to neutralize the analyte.

摘要翻译： 在识别复合体系中，包含随机DNA序列的核酸配体可操作地偶联到有机半导体上，并分布以形成识别复合物的阵列。 当将未知的化学或生物分析物施加到阵列时，一个或多个识别复合物的电和/或光化学性质在核酸配体与分析物结合时被改变。 电和/或光化学性质变化的程度是核酸配体序列对分析物的亲和力的函数。 与阵列相关联的电学和光化学变化作为一个整体，可以用作识别分析物的独特标记。 在某些实施方案中，结合分析物的核酸配体的选择和扩增的迭代过程可用于产生对靶分析物具有更大亲和性和特异性的新阵列，或产生具有高结合力的一个或多个核酸配体 对分析物的亲和力。 本发明还提供了制备以高亲和力结合分析物并使用这种核酸配体中和分析物的核酸配体的方法。

公开(公告)号：US5464768A

公开(公告)日：1995-11-07

申请号：US243350

申请日：1994-05-16

申请人： Johnathan L. Kiel ,  Jill E. Parker ,  John G. Bruno

发明人： Johnathan L. Kiel ,  Jill E. Parker ,  John G. Bruno

IPC分类号： C12N9/02 ,  C12N15/85

CPC分类号： C12N9/0036

摘要： Mamammalian cell lines capable of enhanced nitrite production are by transfecting a murine macrophage or murine thymoma with barley nitrate reductase gene (NR).

摘要翻译： 通过用大麦硝酸还原酶基因（NR）转染鼠巨噬细胞或鼠胸腺瘤，能够增强亚硝酸盐产生的Mamammalian细胞系。

公开(公告)号：US20140011200A1

公开(公告)日：2014-01-09

申请号：US12378933

申请日：2009-02-20

申请人： John G. Bruno ,  Joseph Chanpong

发明人： John G. Bruno ,  Joseph Chanpong

IPC分类号： G01N21/64

CPC分类号： G01N21/6486 ,  C12N15/111 ,  C12N2310/16 ,  C12N2320/10 ,  C12Q1/6804 ,  G01N33/5308 ,  G01N33/542 ,  C12Q2565/101 ,  C12Q2537/161 ,  C12Q2525/205

摘要： Methods are described for the production and use of fluorescence resonance energy transfer (FRET)-based competitive displacement aptamer assay formats. The assay schemes involve FRET in which the analyte (target) is quencher (Q)-labeled and previously bound by a fluorophore (F)-labeled aptamer such that when unlabeled analyte is added to the system and excited by specific wavelengths of light, the fluorescence intensity of the system changes in proportion to the amount of unlabeled analyte added. Alternatively, the aptamer can be Q-labeled and previously bound to an F-labeled analyte so that when unlabeled analyte enters the system, the fluorescence intensity also changes in proportion to the amount of unlabeled analyte. The F or Q is covalently linked to nucleotide triphosphates (NTPs), which are incorporated into the aptamer by various nucleic acid polymerases, such as Taq or Deep Vent Ex& during PCR or asymmetric PCR, and then selected by affinity chromatography, size-exclusion, and fluorescence techniques.

摘要翻译： 描述了用于生产和使用基于荧光共振能量转移（FRET）的竞争性置换适配体测定形式的方法。 测定方案涉及FRET，其中分析物（靶）被猝灭剂（Q）标记并预先通过荧光团（F）标记的适体结合，使得当将未标记的分析物加入到体系中并被特定波长的光激发时， 系统的荧光强度与未标记的分析物的添加量成比例地变化。 或者，适体可以被Q标记并且​​预先与F标记的分析物结合，使得当未标记的分析物进入系统时，荧光强度也与未标记的分析物的量成比例地变化。 F或Q与核苷三磷酸（NTPs）共价连接，其通过各种核酸聚合酶例如Taq或Deep Vent Ex＆在PCR或不对称PCR中并入适配体中，然后通过亲和层析，大小排阻， 和荧光技术。

公开(公告)号：US08389710B2

公开(公告)日：2013-03-05

申请号：US12716088

申请日：2010-03-02

申请人： John G. Bruno ,  Judson C. Miner

发明人： John G. Bruno ,  Judson C. Miner

IPC分类号： C07H21/04

CPC分类号： C07K14/00 ,  A61K47/549 ,  A61K47/64 ,  A61K47/643 ,  A61K47/645 ,  A61K47/6835 ,  C12N15/111 ,  C12N15/115 ,  C12N2310/16 ,  C12N2310/3513 ,  C12N2320/30 ,  C12N2320/51

摘要： Methods are described for improvement of the serum half life of therapeutic nucleic acids by 3′ conjugation to useful target proteins, or other large molecules with useful function. In one embodiment, a 3′ A, C or G overhang is added to ds-DNA and the primary amines conjugated using biocompatible bifunctional linkers to proteins. The resulting nucleic acid-3′ conjugates are serum nuclease-resistant and retained in vivo for long periods without rapid kidney clearance. Further, the choice of conjugate imparts additional functionality to the nucleic acid-3′ conjugate.

摘要翻译： 描述了通过3'缀合到有用靶蛋白或具有有用功能的其它大分子来改善治疗性核酸的血清半衰期的方法。 在一个实施方案中，将3'A，C或G突出端加入到ds-DNA中，并且使用生物相容性双功能接头与蛋白质缀合的伯胺。 所得到的核酸-3'缀合物是血清核酸酶抗性的并且长时间保留在体内而没有快速的肾清除。 此外，缀合物的选择赋予核酸-3'缀合物附加的功能。

公开(公告)号：US5976887A

公开(公告)日：1999-11-02

申请号：US866571

申请日：1997-06-02

申请人： John G. Bruno ,  Jimmy C. Cornette

发明人： John G. Bruno ,  Jimmy C. Cornette

IPC分类号： C09K11/06 ,  C09K11/07 ,  C09K15/32 ,  F21K2/06 ,  G01N33/18 ,  G01N33/20 ,  G01N33/22 ,  G01N33/24

CPC分类号： G01N33/24 ,  C09K11/06 ,  C09K11/07 ,  F21K2/06 ,  G01N33/18 ,  C09K2211/1007 ,  C09K2211/1011 ,  C09K2211/1029 ,  C09K2211/185 ,  C09K2211/188 ,  Y10T436/17 ,  Y10T436/173845

摘要： A novel electrochemiluminescent (ECL) reaction between diaminoaromatic ligands and soluble metal ions, specifically reactions between aminoaromatic ligands, such as 2,4-diaminotoluene (2,4,DAT), 3,4-diaminotoluene (3,4,DAT) and 2,3-diaminonaphthalene (2,3-DAN) and metal ions such as Au(I), Cu(II), Cr(VI), Fe(III), Ru)III), Se(IV) and V(V). Such reactions form the basis for ECL assays in detection of various substances, such as the reactants. The ECL interaction between these substances can also form the basis for binding methods in the detection of other substances, such as nucleic acids and antibodies wherein the metal ion ligand ECL complex may be used as a label. The ECL assays are considered useful for carrying out field and laboratory analyses for the detection of TNT breakdown products and toxic metals in wastewater streams, soil, and ground water supplies. In view of the formation of such ECL complexes being dependent on molecular size, further uses are contemplated for measuring atomic size or intermolecular distances of the complexes formed.

摘要翻译： 二氨基芳族配体和可溶性金属离子之间的新型电化学发光（ECL）反应，特别是氨基芳族配体如2,4-二氨基甲苯（2,4，DAT），3,4-二氨基甲苯（3,4，DAT）和2 （II），Cr（VI），Fe（III），Ru）III），Se（IV）和V（V）等金属离子， 。 这些反应形成了用于检测各种物质如反应物的ECL测定的基础。 这些物质之间的ECL相互作用也可以形成检测其它物质（例如其中金属离子配体ECL复合物可用作标记物的核酸和抗体）的结合方法的基础。 ECL测定被认为可用于对废水，土壤和地下水供应中TNT分解产物和有毒金属的检测进行现场和实验室分析。 考虑到依赖于分子尺寸的这种ECL复合物的形成，考虑到形成的络合物的原子尺寸或分子间距离的进一步用途。

公开(公告)号：US20120270221A1

公开(公告)日：2012-10-25

申请号：US12931305

申请日：2011-01-28

申请人： John G. Bruno ,  Joseph Chanpong

发明人： John G. Bruno ,  Joseph Chanpong

IPC分类号： G01N21/64 ,  B82Y15/00

CPC分类号： C12N15/111 ,  C12N2310/16 ,  C12N2320/10 ,  G01N33/5308 ,  G01N33/533 ,  G01N33/542

摘要： The present invention describes methods for the production and selecting of single chain (single-stranded) fluorescence resonance energy transfer (“FRET”) DNA or RNA aptamers containing fluorophores (F) and quenchers (Q) at various loci within their structures, such that when its specific matching analyte is bound and the FRET-aptamers are excited by specific wavelengths of light, the fluorescence intensity of the system is modulated (increased or decreased) in proportion to the amount of analyte added. F and Q are covalently linked to nucleotide triphosphates (NTPs), which are incorporated by various nucleic acid polymerases such as Taq polymerase during the polymerase chain reaction (PCR) and then selected by affinity chromatographic, size-exclusion or molecular sieving, and fluorescence techniques. Further separation of related FRET-aptamers can be achieved by ion-pair reverse phase high performance liquid chromatography (HPLC) or other types of chromatography. Finally, FRET-aptamer structures and the specific locations of F and Q within FRET-aptamer structures are determined by digestion with exonucleases and mass spectral nucleotide sequencing analysis. Alternatively, single DNA or RNA intrachain FRET-aptamers can be sequenced and the locations of F and Q within the structure can be determined by nanopore sequencing and the locations of F and Q within the structure can be verified by nucleic acid “combing” coupled to high-powered fluorescence microscopy.

摘要翻译： 本发明描述了在其结构内的各个位点生产和选择含有荧光团（F）和猝灭剂（Q）的单链（单链）荧光共振能量转移（FRET）DNA或RNA适体，使得当其 特异性匹配分析物结合，FRET适体被特定波长的光激发，系统的荧光强度与添加的分析物的量成比例地调节（增加或减少）。 F和Q与核苷酸三磷酸（NTPs）共价连接，其在聚合酶链反应（PCR）期间由各种核酸聚合酶例如Taq聚合酶掺入，然后通过亲和层析，大小排阻或分子筛选和荧光技术 。 通过离子对反相高效液相色谱（HPLC）或其他色谱法可以进一步分离相关的FRET-适体。 最后，通过用核酸外切酶消化和质谱核苷酸测序分析来测定FRET-适体结构中F和Q在FRET-适体结构中的特异性位置。 或者，可以对单个DNA或RNA链内FRET-适体进行测序，并且可以通过纳米孔测序确定结构内F和Q的位置，并且结构内的F和Q的位置可以通过核酸结合到高效液相色谱 动力荧光显微镜。

公开(公告)号：US20120135540A1

公开(公告)日：2012-05-31

申请号：US13199484

申请日：2011-08-31

申请人： John G. Bruno

发明人： John G. Bruno

IPC分类号： G01N33/53 ,  C07H21/04

CPC分类号： C12N15/115 ,  G01N33/5308 ,  G01N33/66 ,  G01N33/6887 ,  G01N33/82 ,  G01N2333/4712 ,  G01N2333/4737 ,  G01N2333/54 ,  G01N2333/5412 ,  G01N2333/61

摘要： Specific DNA sequences for binding various clinically relevant analytes from the human body are described. Each of these sequences or their linear, two- and three-dimensional linked sequences can function in varying assay and sensor formats with varying degrees of success. Linkage of the whole or partial DNA sequences (putative binding sites) can be used to enhance specificity and affinity towards complex targets, thereby improving assay selectivity and sensitivity in many instances. In addition, a FRET-based quantitative method is described for normalizing analyte data by assessing urine creatinine and urea levels. Finally, a method is described for removing creatinine or urea by size-exclusion chromatography prior to a FRET-based aptamer assay to avoid the denaturing effects of these compounds.

摘要翻译： 描述了用于结合来自人体的各种临床相关分析物的特异性DNA序列。 这些序列中的每一个或其线性，二维和三维连接的序列可以以不同的测定和传感器形式发挥不同程度的成功。 可以使用全部或部分DNA序列（推定的结合位点）的连接来增强对复杂靶标的特异性和亲和力，从而在许多情况下提高测定选择性和灵敏度。 此外，描述了基于FRET的定量方法，用于通过评估尿肌酐和尿素水平来归一化分析物数据。 最后，描述了在基于FRET的适体测定之前通过尺寸排阻色谱法除去肌酸酐或尿素的方法，以避免这些化合物的变性作用。

公开(公告)号：US20120123096A1

公开(公告)日：2012-05-17

申请号：US12716088

申请日：2010-03-02

申请人： John G. Bruno ,  Judson C. Miner

发明人： John G. Bruno ,  Judson C. Miner

IPC分类号： C07K14/435 ,  C07K14/765 ,  C07K14/745 ,  C07K14/79

CPC分类号： C07K14/00 ,  A61K47/549 ,  A61K47/64 ,  A61K47/643 ,  A61K47/645 ,  A61K47/6835 ,  C12N15/111 ,  C12N15/115 ,  C12N2310/16 ,  C12N2310/3513 ,  C12N2320/30 ,  C12N2320/51

摘要： Methods are described for improvement of the serum half life of therapeutic nucleic acids by 3′ conjugation to useful target proteins, or other large molecules with useful function. In one embodiment, a 3′ A, C or G overhang is added to ds-DNA and the primary amines conjugated using biocompatible bifunctional linkers to proteins. The resulting nucleic acid-3′ conjugates are serum nuclease-resistant and retained in vivo for long periods without rapid kidney clearance. Further, the choice of conjugate imparts additional functionality to the nucleic acid-3′ conjugate.

摘要翻译： 描述了通过3'缀合到有用靶蛋白或具有有用功能的其它大分子来改善治疗性核酸的血清半衰期的方法。 在一个实施方案中，将3'A，C或G突出端加入到ds-DNA中，并且使用生物相容性双功能接头与蛋白质缀合的伯胺。 所得到的核酸-3'缀合物是血清核酸酶抗性的并且长时间保留在体内而没有快速的肾清除。 此外，缀合物的选择赋予核酸-3'缀合物附加的功能。

公开(公告)号：US09562900B2

公开(公告)日：2017-02-07

申请号：US13136820

申请日：2011-08-11

申请人： John G. Bruno

发明人： John G. Bruno

IPC分类号： C07H21/04 ,  G01N33/569 ,  C12N15/115 ,  C12Q1/68

CPC分类号： G01N33/56916 ,  C12N15/115 ,  C12N2310/16 ,  C12Q1/689 ,  G01N33/56922 ,  Y02A50/451

摘要： Specific DNA sequences for binding various foodborne and waterborne pathogens and biotoxins are described. Each of these sequences can function in varying assay and sensor formats with varying degrees of success.

摘要翻译： 描述了用于结合各种食源性和水源性病原体和生物毒素的特异性DNA序列。 这些序列中的每一个可以以不同的测定和传感器形式起作用，具有不同程度的成功。