公开(公告)号：US10072057B2

公开(公告)日：2018-09-11

申请号：US15656740

申请日：2017-07-21

申请人： ModernaTX, Inc.

发明人： Stephen G. Hoge ,  William Joseph Issa ,  Edward John Miracco ,  Jennifer Nelson ,  John Reynders ,  Matthew Stanton

IPC分类号： C07K14/535 ,  C12P19/34 ,  C07K14/505 ,  C12N9/00 ,  A61K48/00 ,  C12P21/02 ,  C12N9/02

CPC分类号： C07K14/535 ,  A61K48/005 ,  C07K14/505 ,  C12N9/0069 ,  C12P19/34 ,  C12P21/02 ,  C12Y113/12007

摘要： The present disclosure provides alternative nucleosides, nucleotides, and nucleic acids, and methods of using them. In some aspects, the disclosure provides mRNA wherein the uracil content has been modified and which may be particularly effective for use in therapeutic compositions, because they may benefit from both high expression levels and limited induction of the innate immune response. In some aspects, the disclosure provides methods for the production of pharmaceutical compositions including mRNA without reverse phase chromatography.

公开(公告)号：US11377470B2

公开(公告)日：2022-07-05

申请号：US14776864

申请日：2014-03-13

申请人： ModernaTX, Inc.

发明人： William Joseph Issa ,  John Grant Aunins ,  Stephane Bancel

IPC分类号： C07H21/02 ,  C07H1/06 ,  C08B37/00 ,  C12Q1/6806 ,  C12N15/10

摘要： Disclosed herein are methods for purifying RNA comprising poly A. Also disclosed herein are compositions such as surfaces and oligonucleotides for purifying RNA comprising polyA. Other embodiments are also disclosed. Commercially-available resins having polythymidine oligonucleotide ligands typically contain less than 30 thymidine (2′deoxy) residues and some commercial resin suppliers utilize a distribution of dT chain lengths, not of a discreet length.

公开(公告)号：US11845772B2

公开(公告)日：2023-12-19

申请号：US17854187

申请日：2022-06-30

申请人： ModernaTX, Inc.

发明人： William Joseph Issa ,  John Grant Aunins ,  Stephane Bancel

IPC分类号： C07H21/02 ,  C12Q1/6806 ,  C12N15/10 ,  C07H1/06 ,  C08B37/00

CPC分类号： C07H21/02 ,  C07H1/06 ,  C08B37/0039 ,  C12N15/1006 ,  C12Q1/6806

摘要： Disclosed herein are methods for purifying RNA comprising poly A. Also disclosed herein are compositions such as surfaces and oligonucleotides for purifying RNA comprising polyA. Other embodiments are also disclosed. Commercially-available resins having polythymidine oligonucleotide ligands typically contain less than 30 thymidine (2′deoxy) residues and some commercial resin suppliers utilize a distribution of dT chain lengths, not of a discreet length.

公开(公告)号：US10858647B2

公开(公告)日：2020-12-08

申请号：US16049132

申请日：2018-07-30

申请人： ModernaTX, Inc.

发明人： William Joseph Issa ,  Yuxun Wang ,  Stephane Bancel

IPC分类号： C12N15/10 ,  C12Q1/6806

摘要： The present invention describes methods of removing DNA from an RNA transcript during the mRNA production process. The method embodies procedures for obtaining an in vitro transcription product, and removing any DNA from the product. The DNA can be removed by adding either free DNase or a resin containing immobilized DNase to the product, and recovering the RNA transcript. Alternatively, the DNA template used in the in vitro transcription reaction is labeled. After transcription, the product is applied to a resin that is configured to bind the label, and the RNA transcript is recovered. To detect whether any residual impurities are left in the RNA transcript product, the product is subjected to nuclease digestion and subsequently to liquid chromatography-tandem mass spectrometry analysis to quantitate any residual DNA. The present invention demonstrates efficient and effective methods of isolating an RNA transcript from an in vitro transcription product.

公开(公告)号：US10138507B2

公开(公告)日：2018-11-27

申请号：US14777190

申请日：2014-03-13

申请人： ModernaTX, Inc.

发明人： Stephane Bancel ,  William Joseph Issa ,  John Grant Aunins ,  Tirtha Chakraborty

IPC分类号： C12P19/34 ,  C12Q1/68 ,  C12N15/10 ,  C12Q1/6865

摘要： Described are methods for production of RNA transcripts using a non-amplified, linearized DNA template in an in vitro transcription reaction. Enzymatic 5′ capping and oligo dT purification can also be included in the methods.

公开(公告)号：US09751925B2

公开(公告)日：2017-09-05

申请号：US14960031

申请日：2015-12-04

申请人： ModernaTX, Inc.

发明人： Stephen G. Hoge ,  William Joseph Issa ,  Edward John Miracco ,  Jennifer Nelson ,  John Reynders ,  Matthew Stanton

IPC分类号： C07K14/535 ,  C12P19/34 ,  C07K14/505 ,  C12N9/00 ,  C12N9/02

CPC分类号： C07K14/535 ,  A61K48/005 ,  C07K14/505 ,  C12N9/0069 ,  C12P19/34 ,  C12P21/02 ,  C12Y113/12007

摘要： The present disclosure provides alternative nucleosides, nucleotides, and nucleic acids, and methods of using them. In some aspects, the disclosure provides mRNA wherein the uracil content has been modified and which may be particularly effective for use in therapeutic compositions, because they may benefit from both high expression levels and limited induction of the innate immune response. In some aspects, the disclosure provides methods for the production of pharmaceutical compositions including mRNA without reverse phase chromatography.

公开(公告)号：US10590161B2

公开(公告)日：2020-03-17

申请号：US14777320

申请日：2014-03-14

申请人： ModernaTX, Inc.

发明人： William Joseph Issa ,  Joseph Louis Barberio ,  John Grant Aunins ,  Noubar B. Afeyan

IPC分类号： C07H21/02 ,  C12N15/10 ,  G01N30/96 ,  B01D15/36 ,  G01N30/88

摘要： The current landscape for preparative chromatographic RNA purification uses reversed phase HPLC, but this technique presents many issues with process scale up and ion exchange for preparative purification has only been used for short RNAs. The invention provides preparative purification of RNA (e.g., mRNA) using ion (e.g., anion) exchange chromatography that allows for separation of longer RNAs up to 10,000 nucleotides in length via a scalable method. This method avoids problems with current techniques by using low pressure chromatography that is agreeable with existing equipment in cGMP commercial facilities, that uses aqueous-bases solutions as the mobile phase (rather than flammable of greater than 10 mg RNA/mL resin (e.g., using larger pore sorbents, >500 Angstroms, that display greater mRNA binding capacities), and that yields desired RNA salt forms for downstream formulation with no additional manipulation necessary (unlike ion pair reverse phase techniques).

公开(公告)号：US10077439B2

公开(公告)日：2018-09-18

申请号：US14777301

申请日：2014-03-13

申请人： ModernaTX, Inc.

发明人： William Joseph Issa ,  Yuxun Wang ,  Stephane Bancel

IPC分类号： C12Q1/68 ,  C07H21/00 ,  C12N15/10 ,  C12Q1/6806

CPC分类号： C12N15/1006 ,  C12N15/101 ,  C12Q1/6806 ,  C12Q2521/319 ,  C12Q2565/137

摘要： The present invention describes methods of removing DNA from an RNA transcript during the mRNA production process. The method embodies procedures for obtaining an in vitro transcription product, and removing any DNA from the product. The DNA can be removed by adding either free DNase or a resin containing immobilized DNase to the product, and recovering the RNA transcript. Alternatively, the DNA template used in the in vitro transcription reaction is labeled. After transcription, the product is applied to a resin that is configured to bind the label, and the RNA transcript is recovered. To detect whether any residual impurities are left in the RNA transcript product, the product is subjected to nuclease digestion and subsequently to liquid chromatography-tandem mass spectrometry analysis to quantitate any residual DNA. The present invention demonstrates efficient and effective methods of isolating an RNA transcript from an in vitro transcription product.

公开(公告)号：US20180009866A1

公开(公告)日：2018-01-11

申请号：US15656740

申请日：2017-07-21

申请人： ModernaTX, Inc.

发明人： Stephen G. Hoge ,  William Joseph Issa ,  Edward John Miracco ,  Jennifer Nelson ,  John Reynders ,  Matthew Stanton

IPC分类号： C07K14/535 ,  C07K14/505 ,  C12N9/02 ,  C12P19/34

CPC分类号： C07K14/535 ,  A61K48/005 ,  C07K14/505 ,  C12N9/0069 ,  C12P19/34 ,  C12P21/02 ,  C12Y113/12007

摘要： The present disclosure provides alternative nucleosides, nucleotides, and nucleic acids, and methods of using them. In some aspects, the disclosure provides mRNA wherein the uracil content has been modified and which may be particularly effective for use in therapeutic compositions, because they may benefit from both high expression levels and limited induction of the innate immune response. In some aspects, the disclosure provides methods for the production of pharmaceutical compositions including mRNA without reverse phase chromatography.