Patents by Inventor Mark Ambroso
Mark Ambroso has filed for patents to protect the following inventions. This listing includes patent applications that are pending as well as patents that have already been granted by the United States Patent and Trademark Office (USPTO).
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Publication number: 20240117428Abstract: The present disclosure provides nucleotide conjugates each configured to include a core attached to multiple nucleotide-arms, where the nucleotide-arms are modular and comprise (i) a core attachment moiety, (ii) a spacer, (iii) a linker, and (iv) a nucleotide unit. The nucleotide unit of each nucleotide-arm can bind a polymerase which is complexed with a nucleic acid template and nucleic acid primer. The nucleotide unit can bind the 3? end of the primer at a position that is opposite a complementary nucleotide in the template strand. Under suitable conditions, the nucleotide unit of the nucleotide conjugates binds the primer strand but does not undergo polymerase-catalyzed incorporation. The binding event can be detected, and the specific base of the nucleotide unit can be identified. The nucleotide conjugates described herein are useful for nucleic acid sequencing methods, particularly for massively parallel sequencing methods employed for next gen sequencing platforms.Type: ApplicationFiled: August 2, 2023Publication date: April 11, 2024Inventors: MICHAEL PREVITE, MARK AMBROSO, TYLER LOPEZ, MICHAEL KLEIN, VIRGINIA SAADE, MATTHEW KELLINGER
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Patent number: 11859241Abstract: The present disclosure provides compositions and methods that employ the compositions for conducting pairwise sequencing and for generating concatemer template molecules for pairwise sequencing. The concatemers can be generated using a rolling circle amplification reaction which is conducted either on-support, or conducted in-solution and then distributed onto a support. The rolling circle amplification reaction generates concatemers containing tandem copies of a sequence of interest and at least one universal adaptor sequence. An increase in the number of tandem copies in a given concatemer increases the number of sites along the concatemer for hybridizing to multiple sequencing primers which serve as multiple initiation sites for polymerase-catalyzed sequencing reactions. When the sequencing reaction employs detectably labeled nucleotides and/or detectably labeled multivalent molecules (e.g.Type: GrantFiled: February 8, 2023Date of Patent: January 2, 2024Assignee: Element Biosciences, Inc.Inventors: Sinan Arslan, Junhua Zhao, Molly He, Samantha Snow, William Light, Matthew Kellinger, Michael Previte, Michael Kim, Hua Yu, Yu-Hsien Hwang-Fu, Marco Tjioe, Andrew Boddicker, Mark Ambroso, Tyler Lopez, Michael Klein, Virginia Saade
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Publication number: 20230332222Abstract: Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L135I in the mature alpha-hemolysin amino acid sequence. The ?-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.Type: ApplicationFiled: February 21, 2023Publication date: October 19, 2023Applicant: Roche Sequencing Solutions, Inc.Inventors: Mark Ambroso, Timothy Craig, Matthew DiPietro, Corissa Harris, Marshall Porter
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Patent number: 11788075Abstract: Provided herein are engineered variants of archaeal polymerases that exhibit exonuclease-minus activity, enhanced thermostability, enhanced incorporation of 3? modified nucleotides, improved uracil-tolerance and/or reduce sequence-specific errors in polymerase-catalyzed nucleotide binding and extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases and forming binding complexes, and uses for conducting nucleic acid sequencing reactions.Type: GrantFiled: January 27, 2023Date of Patent: October 17, 2023Assignee: ELEMENT BIOSCIENCES, INC.Inventors: Jendrik Hentschel, Tyler Lopez, Michael Klein, Virginia Saade, Matthew Kellinger, Mark Ambroso
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Publication number: 20230265400Abstract: Provided herein are engineered variants of archaeal polymerases that exhibit exonuclease-minus activity, enhanced thermostability, enhanced incorporation of 3? modified nucleotides, improved uracil-tolerance and/or reduce sequence-specific errors in polymerase-catalyzed nucleotide binding and extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases and forming binding complexes, and uses for conducting nucleic acid sequencing reactions.Type: ApplicationFiled: January 27, 2023Publication date: August 24, 2023Inventors: Jendrik HENTSHCEL, Tyler LOPEZ, Michael KLEIN, Virginia SAADE, Matthew KELLINGER, Mark AMBROSO
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Publication number: 20230265401Abstract: Provided herein are engineered variants of archaeal polymerases that exhibit exonuclease-minus activity, enhanced thermostability, enhanced incorporation of 3? modified nucleotides, improved uracil-tolerance and/or reduce sequence-specific errors in polymerase-catalyzed nucleotide binding and extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases and forming binding complexes, and uses for conducting nucleic acid sequencing reactions.Type: ApplicationFiled: January 27, 2023Publication date: August 24, 2023Inventors: Jendrik HENTSHCEL, Tyler LOPEZ, Michael KLEIN, Virginia SAADE, Matthew KELLINGER, Mark AMBROSO
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Publication number: 20230265402Abstract: Provided herein are engineered variants of archaeal polymerases that exhibit exonuclease-minus activity, enhanced thermostability, enhanced incorporation of 3? modified nucleotides, improved uracil-tolerance and/or reduce sequence-specific errors in polymerase-catalyzed nucleotide binding and extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases and forming binding complexes, and uses for conducting nucleic acid sequencing reactions.Type: ApplicationFiled: January 27, 2023Publication date: August 24, 2023Inventors: Jendrik HENTSHCEL, Tyler LOPEZ, Michael KLEIN, Virginia SAADE, Matthew KELLINGER, Mark AMBROSO
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Publication number: 20230203564Abstract: The present disclosure provides compositions and methods that employ the compositions for conducting pairwise sequencing and for generating concatemer template molecules for pairwise sequencing. The concatemers can be generated using a rolling circle amplification reaction which is conducted either on-support, or conducted in-solution and then distributed onto a support. The rolling circle amplification reaction generates concatemers containing tandem copies of a sequence of interest and at least one universal adaptor sequence. An increase in the number of tandem copies in a given concatemer increases the number of sites along the concatemer for hybridizing to multiple sequencing primers which serve as multiple initiation sites for polymerase-catalyzed sequencing reactions. When the sequencing reaction employs detectably labeled nucleotides and/or detectably labeled multivalent molecules (e.g.Type: ApplicationFiled: February 8, 2023Publication date: June 29, 2023Inventors: Sinan ARSLAN, Junhua ZHAO, Molly HE, Samantha SNOW, William LIGHT, Matthew KELLINGER, Michael PREVITE, Michael KIM, Hua YU, Yu-Hsien HWANG-FU, Marco TJIOE, Andrew BODDICKER, Mark AMBROSO, Tyler LOPEZ, Michael KLEIN, Virginia SAADE
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Patent number: 11613778Abstract: Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L135I in the mature alpha-hemolysin amino acid sequence. The ?-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.Type: GrantFiled: January 25, 2021Date of Patent: March 28, 2023Assignee: Roche Sequencing Solutions, Inc.Inventors: Mark Ambroso, Timothy Craig, Matthew DiPietro, Corissa Harris, Marshall Porter
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Publication number: 20220403351Abstract: Provided herein are engineered variants of archaeal, prokaryotic, and eukaryotic polymerases that exhibit enhanced thermostability, enhanced incorporation of 3? modified nucleotides, and improved uracil-tolerance, in polymerase-catalyzed nucleotide extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases, forming binding complexes and forming ternary complexes, and uses for conducting nucleic acid sequencing reactions.Type: ApplicationFiled: March 25, 2022Publication date: December 22, 2022Inventors: Mark Ambroso, Tyler Lopez, Michael Klein, Virginia Saade, Matthew Kellinger
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Publication number: 20220403353Abstract: Provided herein are engineered variants of archaeal, prokaryotic, and eukaryotic polymerases that exhibit enhanced thermostability, enhanced incorporation of 3? modified nucleotides, and improved uracil-tolerance, in polymerase-catalyzed nucleotide extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases, forming binding complexes and forming ternary complexes, and uses for conducting nucleic acid sequencing reactions.Type: ApplicationFiled: March 25, 2022Publication date: December 22, 2022Inventors: Mark Ambroso, Tyler Lopez, Michael Klein, Virginia Saade, Matthew Kellinger
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Publication number: 20220403352Abstract: Provided herein are engineered variants of archaeal, prokaryotic, and eukaryotic polymerases that exhibit enhanced thermostability, enhanced incorporation of 3? modified nucleotides, and improved uracil-tolerance, in polymerase-catalyzed nucleotide extension reactions relative to wild type polymerase enzymes. Also provided are uses of the engineered polymerases for forming complexed polymerases, forming binding complexes and forming ternary complexes, and uses for conducting nucleic acid sequencing reactions.Type: ApplicationFiled: March 25, 2022Publication date: December 22, 2022Inventors: Mark Ambroso, Tyler Lopez, Michael Klein, Virginia Saade, Matthew Kellinger
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Patent number: 11150233Abstract: Described herein are nanopore protein conjugates that can be used in DNA sequencing reactions. The nanopore protein conjugates includes a nanopore protein monomer that is joined to a DNA binding domain. The nanopore protein monomer is available to oligomerize with other nanopore protein monomers, while the DNA binding domain is available to bind to a template DNA strand. In certain examples, the nanopore protein monomer is an alpha-hemolysin monomer or variant thereof and the DNA binding domain is an Sso7d protein or variant thereof, such as an Sso7d-like protein. Also provided are nanopore protein assemblies incorporating the nanopore protein conjugates, along with methods of using the nanopore protein assemblies in sequencing reactions.Type: GrantFiled: September 28, 2018Date of Patent: October 19, 2021Assignee: Roche Sequencing Solutions, Inc.Inventors: Timothy Craig, Corissa Harris, Matt Dipetro, Mark Ambroso, Yaozhong Zou, Marshall Porter, Issa Delkaninia, Alexxa Noble, James Fairman, Wiseley Wu, Kapil Bajaj, Giovanni Bellesia, Seong-Ho Shin
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Publication number: 20210230683Abstract: Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L1351 in the mature alpha-hemolysin amino acid sequence. The ?-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.Type: ApplicationFiled: January 25, 2021Publication date: July 29, 2021Applicant: Roche Sequencing Solutions, Inc.Inventors: Mark Ambroso, Timothy Craig, Matthew DiPietro, Corissa Harris, Marshall Porter
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Patent number: 10934582Abstract: Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L1351 in the mature alpha-hemolysin amino acid sequence. The ?-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.Type: GrantFiled: July 23, 2019Date of Patent: March 2, 2021Assignee: Roche Sequencing Solutions, Inc.Inventors: Mark Ambroso, Timothy Craig, Matthew DiPietro, Corissa Harris, Marshall Porter
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Publication number: 20190376134Abstract: Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L1351 in the mature alpha-hemolysin amino acid sequence. The a-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.Type: ApplicationFiled: July 23, 2019Publication date: December 12, 2019Applicant: Roche Sequencing Solutions, Inc.Inventors: Mark Ambroso, Timothy Craig, Matthew DePietro, Corissa Harris, Marshall Porter
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Publication number: 20190079067Abstract: Described herein are nanopore protein conjugates that can be used in DNA sequencing reactions. The nanopore protein conjugates includes a nanopore protein monomer that is joined to a DNA binding domain. The nanopore protein monomer is available to oligomerize with other nanopore protein monomers, while the DNA binding domain is available to bind to a template DNA strand. In certain examples, the nanopore protein monomer is an alpha-hemolysin monomer or variant thereof and the DNA binding domain is an Sso7d protein or variant thereof, such as an Sso7d-like protein. Also provided are nanopore protein assemblies incorporating the nanopore protein conjugates, along with methods of using the nanopore protein assemblies in sequencing reactions.Type: ApplicationFiled: September 28, 2018Publication date: March 14, 2019Applicant: GENIA TECHNOLOGIES, INC.Inventors: Timothy Craig, Corissa Harris, Matt Dipetro, Mark Ambroso, Yaozhong Zou, Marshall Porter, Issa Delkaninia, Alexxa Noble, James Fairman, Wiseley Wu, Kapil Bajaj, Giovanni Bellesia, Seong-Ho Shin
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Publication number: 20180002750Abstract: Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and/or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L135I in the mature alpha-hemolysin amino acid sequence. The ?-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.Type: ApplicationFiled: June 29, 2017Publication date: January 4, 2018Inventors: Mark Ambroso, Timothy Craig, Matthew DiPietro, Corissa Harris, Marshall Porter