公开(公告)号：US20140308730A1

公开(公告)日：2014-10-16

申请号：US14087307

申请日：2013-11-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Theo NIKIFOROV ,  Daniel MAZUR ,  Xinzhan PENG ,  Tommie Lloyd LINCECUM ,  Yuri BELOSLUDTSEV ,  Howard REESE ,  Dmitriy GREMYACHINSKIY ,  Roman ROZHKOV ,  John MAURO ,  Joseph BEECHEM ,  Eric TULSKY ,  Imad NAASANI ,  Kari HALEY ,  Joseph TREADWAY

IPC: C12N9/12

CPC classification number: C12Q1/6869 ,  C07H19/20 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6818 ,  C12Y207/07 ,  C12Y207/07007 ,  G01N21/6428 ,  G01N33/582 ,  G01N2021/6432

Abstract: Disclosed herein are conjugates comprising a biomolecule linked to a label that have biological activity and are useful in a wide variety of biological applications. For example, provided herein are polymerase-nanoparticle conjugates including a polymerase linked to a nanoparticle, wherein the conjugate has polymerase activity. Such conjugates can exhibit reduced aggregation and improved stochiometries wherein the average biomolecule:nanoparticle ratio approaches or equals 1:1. Also disclosed herein are improved methods for preparing such conjugates, and methods and systems for using such conjugates in biological applications such as nucleotide incorporation, primer extension and single molecule sequencing.

Abstract translation: 本文公开了包含连接到具有生物活性并可用于多种生物应用的标签的生物分子的缀合物。 例如，本文提供了聚合酶 - 纳米颗粒缀合物，其包括与纳米颗粒连接的聚合酶，其中缀合物具有聚合酶活性。 这样的缀合物可以表现出减少的聚集和改进的沉淀物，其中平均生物分子：纳米颗粒比接近或等于1:1。 本文还公开了制备这种缀合物的改进方法，以及在生物应用中使用这种缀合物的方法和系统，例如核苷酸掺入，引物延伸和单分子测序。

公开(公告)号：US20150118679A1

公开(公告)日：2015-04-30

申请号：US14584829

申请日：2014-12-29

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Joseph BEECHEM ,  Theo NIKIFOROV ,  Vi-En CHOONG ,  Xinzhan PENG ,  Guobin LUO ,  Cheng-Yao CHEN ,  Michael PREVITE

IPC: C12Q1/68

CPC classification number: C12Q1/6869 ,  C07H19/20 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6818 ,  C12Y207/07 ,  C12Y207/07007 ,  G01N21/6428 ,  G01N33/582 ,  G01N2021/6432

Abstract: Provided herein are systems and methods for nucleotide incorporation reactions. The systems comprise polymerases having altered nucleotide incorporation kinetics and are linked to an energy transfer donor moiety, and nucleotide molecules linked with at least one energy transfer acceptor moiety. The donor and acceptor moieties undergo energy transfer when the polymerase and nucleotide are proximal to each other during nucleotide binding and/or nucleotide incorporation. As the donor and acceptor moieties undergo energy transfer, they generate an energy transfer signal which can be associated with nucleotide binding or incorporation. Detecting a time sequence of the generated signals, or the change in the signals, can be used to determine the order of the incorporated nucleotides, and can therefore be used to deduce the sequence of the target molecule.

Abstract translation: 本文提供了用于核苷酸掺入反应的系统和方法。 该系统包含具有改变的核苷酸掺入动力学并且与能量转移供体部分连接的聚合酶，以及与至少一个能量转移受体部分连接的核苷酸分子。 当核苷酸结合和/或核苷酸掺入期间聚合酶和核苷酸彼此接近时，供体和受体部分进行能量转移。 当供体和受体部分进行能量转移时，它们产生可与核苷酸结合或掺入相关联的能量转移信号。 检测所产生的信号的时间序列或信号的变化可用于确定掺入的核苷酸的顺序，因此可用于推导靶分子的序列。

公开(公告)号：US20190062830A1

公开(公告)日：2019-02-28

申请号：US16123166

申请日：2018-09-06

Applicant: Life Technologies Corporation

Inventor： Theo NIKIFOROV ,  Daniel MAZUR ,  Xinzhan PENG ,  Tommie Lloyd LINCECUM ,  Yuri BELOSLUDTSEV ,  Howard REESE ,  Dmitriy GREMYACHINSKIY ,  Roman ROZHKOV ,  John MAURO ,  Joseph BEECHEM ,  Eric TULSKY ,  Imad NAASANI ,  Kari HALEY ,  Joseph TREADWAY

IPC: C12Q1/6869 ,  C12N9/12 ,  G01N33/58 ,  G01N21/64 ,  C07H19/20 ,  C12Q1/6818 ,  C12N9/96

Abstract: Disclosed herein are conjugates comprising a biomolecule linked to a label that have biological activity and are useful in a wide variety of biological applications. For example, provided herein are polymerase-nanoparticle conjugates including a polymerase linked to a nanoparticle, wherein the conjugate has polymerase activity. Such conjugates can exhibit reduced aggregation and improved stochiometries wherein the average biomolecule:nanoparticle ratio approaches or equals 1:1. Also disclosed herein are improved methods for preparing such conjugates, and methods and systems for using such conjugates in biological applications such as nucleotide incorporation, primer extension and single molecule sequencing.

公开(公告)号：US20190062829A1

公开(公告)日：2019-02-28

申请号：US16123140

申请日：2018-09-06

Applicant: Life Technologies Corporation

Inventor： Joseph BEECHEM ,  Theo NIKIFOROV ,  Vi-En CHOONG ,  Xinzhan PENG ,  Guobin LUO ,  Cheng-Yao CHEN ,  Michael PREVITE

IPC: C12Q1/6869 ,  C12N9/12 ,  G01N33/58 ,  G01N21/64 ,  C07H19/20 ,  C12Q1/6818 ,  C12N9/96

Abstract: Provided herein are systems and methods for nucleotide incorporation reactions. The systems comprise polymerases having altered nucleotide incorporation kinetics and are linked to an energy transfer donor moiety, and nucleotide molecules linked with at least one energy transfer acceptor moiety. The donor and acceptor moieties undergo energy transfer when the polymerase and nucleotide are proximal to each other during nucleotide binding and/or nucleotide incorporation. As the donor and acceptor moieties undergo energy transfer, they generate an energy transfer signal which can be associated with nucleotide binding or incorporation. Detecting a time sequence of the generated signals, or the change in the signals, can be used to determine the order of the incorporated nucleotides, and can therefore be used to deduce the sequence of the target molecule.

公开(公告)号：US20170283868A1

公开(公告)日：2017-10-05

申请号：US15621341

申请日：2017-06-13

Applicant: Life Technologies Corporation

Inventor： Joseph BEECHEM ,  Theo NIKIFOROV ,  Vi-En CHOONG ,  Xinzhan PENG ,  Guobin LUO ,  Cheng-Yao CHEN ,  Michael PREVITE

IPC: C12Q1/68 ,  G01N21/64 ,  C12N9/96 ,  C12N9/12 ,  C07H19/20 ,  G01N33/58

CPC classification number: C12Q1/6869 ,  C07H19/20 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6818 ,  C12Y207/07 ,  C12Y207/07007 ,  G01N21/6428 ,  G01N33/582 ,  G01N2021/6432

Abstract: Provided herein are systems and methods for nucleotide incorporation reactions. The systems comprise polymerases having altered nucleotide incorporation kinetics and are linked to an energy transfer donor moiety, and nucleotide molecules linked with at least one energy transfer acceptor moiety. The donor and acceptor moieties undergo energy transfer when the polymerase and nucleotide are proximal to each other during nucleotide binding and/or nucleotide incorporation. As the donor and acceptor moieties undergo energy transfer, they generate an energy transfer signal which can be associated with nucleotide binding or incorporation. Detecting a time sequence of the generated signals, or the change in the signals, can be used to determine the order of the incorporated nucleotides, and can therefore be used to deduce the sequence of the target molecule.

公开(公告)号：US20160265044A1

公开(公告)日：2016-09-15

申请号：US15166372

申请日：2016-05-27

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： Theo NIKIFOROV ,  Daniel MAZUR ,  Xinzhan PENG ,  Tommie Lloyd LINCECUM ,  Yuri BELOSLUDTSEV ,  Howard REESE ,  Dmitriy GREMYACHINSKIY ,  Roman ROZHKOV ,  John MAURO ,  Joseph BEECHEM ,  Eric TULSKY ,  Imad NAASANI ,  Kari HALEY ,  Joseph TREADWAY

IPC: C12Q1/68 ,  C12N9/12 ,  G01N21/64 ,  C12N9/96

CPC classification number: C12Q1/6869 ,  C07H19/20 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6818 ,  C12Y207/07 ,  C12Y207/07007 ,  G01N21/6428 ,  G01N33/582 ,  G01N2021/6432

Abstract: Disclosed herein are conjugates comprising a biomolecule linked to a label that have biological activity and are useful in a wide variety of biological applications. For example, provided herein are polymerase-nanoparticle conjugates including a polymerase linked to a nanoparticle, wherein the conjugate has polymerase activity. Such conjugates can exhibit reduced aggregation and improved stochiometries wherein the average biomolecule:nanoparticle ratio approaches or equals 1:1. Also disclosed herein are improved methods for preparing such conjugates, and methods and systems for using such conjugates in biological applications such as nucleotide incorporation, primer extension and single molecule sequencing.

Abstract translation: 本文公开了包含连接到具有生物活性并可用于多种生物应用的标签的生物分子的缀合物。 例如，本文提供了聚合酶 - 纳米颗粒缀合物，其包括与纳米颗粒连接的聚合酶，其中缀合物具有聚合酶活性。 这样的缀合物可以表现出减少的聚集和改进的沉淀物，其中平均生物分子：纳米颗粒比接近或等于1:1。 本文还公开了制备这种缀合物的改进方法，以及在生物应用中使用这种缀合物的方法和系统，例如核苷酸掺入，引物延伸和单分子测序。