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`UNITED STATES PATENT AND TRADEMARK OFFICE
`_____________________
`
`BEFORE THE PATENT TRIAL AND APPEAL BOARD
`_____________________
`
`
`MODERNATX, INC.
`Petitioner
`
`v.
`
`CUREVAC AG
`Patent Owner
`
`U.S. Patent No. 8,383,340
`_____________________
`
`Case No. IPR2017-02194
`Patent 8,383,340
`_____________________
`
`SECOND DECLARATION OF PROF. DAVID HORNBY, PH.D.
`
`
`
`
`
`MTX1075
`ModernaTX, Inc. v. CureVac AG
`IPR2017-02194
`
`

`

`
`
`
`I.
`II.
`III.
`IV.
`
`V.
`VI.
`
`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`TABLE OF CONTENTS
`
`Introduction ................................................................................................... 1
`Summary of opinions .................................................................................... 2
`List of documents I considered in formulating my opinions ....................... 4
`State of the art before December 22, 2006 ................................................. 13
`A. Overview of RNA ............................................................................... 13
`B. The field of RNA therapeutics was booming by 2006 ........................ 15
`C. By 2006, Ion-Pair Reversed-Phase High Performance Liquid
`Chromatography
`(IP RP HPLC) methods originally
`developed for DNA purification were successfully being
`adapted for purifying RNA ................................................................. 17
`D. By 2006, a POSA would have been acutely familiar with
`strategies for extracting and
`isolating RNA and for
`preventing RNA degradation by RNases ............................................ 21
`Claim construction ...................................................................................... 23
`Claims 1, 3, 4, 6-19, and 21-26 would have been obvious over the
`combination of Zhang and Lloyd ............................................................... 31
`A. A POSA would have had a reason to use Lloyd’s method to
`purify large quantities of Zhang’s RNA .............................................. 31
`1.
`A POSA would have had a reason to purify Zhang’s
`siRNA
`and
`ribozymes—not
`just
`antisense
`oligonucleotides—on a preparative scale ................................ 31
`A POSA would have had a reason to use Lloyd’s
`method for preparative purification of Zhang’s
`antisense RNA .......................................................................... 33
`(a) A POSA would have understood that Lloyd’s
`method can successfully separate a complex
`mixture of oligonucleotides ........................................... 35
`(b) Lloyd expressly suggested using PLPR-S
`columns for preparative-scale purification of
`RNA and taught that the columns have “high
`capacity”—a feature necessary for preparative
`purification ..................................................................... 40
`The art would not have taught a POSA away
`from using Lloyd’s porous stationary phase for
`a preparative-scale purification of RNA ........................ 43
`
`(c)
`
`2.
`
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
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`
`
`IX.
`
`X.
`
`(d) My own publications would not have taught a
`POSA to use a non-porous stationary phase
`over a porous one for preparative purification
`of RNA by 2006............................................................. 46
`B. A POSA would have had a reasonable expectation of
`success ................................................................................................. 47
`1.
`Lloyd, in view of the high level of skill and
`knowledge in the art, would have given a POSA a
`reasonable expectation of success ............................................ 47
`2. Multiple examples in the field of applying IP RP
`HPLC methods developed for DNA to RNA would
`have given a POSA a reasonable expectation of
`success with Lloyd’s method ................................................... 49
`A POSA’s expectation of success would have
`exceeded an expectation of achieving some level of
`purity and resolution required by the claims ........................... 51
`4. My own publications do not demonstrate that methods
`of purifying DNA cannot be used to purify RNA ................... 52
`Dr. Schwenger’s experiments do not diminish a
`POSA’s reasonable expectations of success ............................ 55
`C. Claims 4, 6, and 7 would have been obvious over the
`combination of Zhang and Lloyd ........................................................ 57
`VII. Claim 2 would have been obvious over the combination of Sullenger
`and Lloyd .................................................................................................... 58
`VIII. Claim 5 would have been obvious over the combination of Zhang,
`Lloyd, and the 2004-2005 Polymer Laboratories Catalog ......................... 59
`Claim 20 would have been obvious over the combination of Zhang,
`Lloyd and Gjerde II .................................................................................... 60
`CureVac’s objective evidence do not support non-obviousness of the
`claimed methods ......................................................................................... 61
`A. Dr. Fotin-Mleczek’s protein expression data do not support
`non-obviousness of the claimed methods ........................................... 62
`1.
`Dr. Fotin-Mleczek did not compare the methods with
`the closest prior art ................................................................... 63
`Protein expression results that Dr. Fotin-Mleczek
`discussed were obtained from methods that are not
`representative of the claims as a whole ................................... 64
`
`3.
`
`5.
`
`2.
`
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`3.
`
`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`The results achieved with the claimed methods are
`due to a feature that was disclosed in the prior art .................. 66
`B. Dr. Thran’s data do not support non-obviousness of the
`claimed methods .................................................................................. 67
`1.
`The results presented by Dr. Thran are not reliable ................. 67
`2.
`Dr. Thran did not compare the results with the closest
`prior art ..................................................................................... 69
`The results achieved with the claimed methods are
`due to a feature that was disclosed in the prior art .................. 69
`The
`results presented by Dr. Thran are not
`representative of the claims as a whole ................................... 70
`The discussion of Dr. Thran’s experiments in Dr.
`Fotin-Mleczek’s declaration suffers from the same
`shortcomings as Dr. Thran’s data ............................................ 71
`Gjerde I anticipates claims 1-5, 8, 10-22, and 26 ....................................... 72
`A. Gjerde I teaches a “porous reverse phase” under either claim
`construction ......................................................................................... 72
`B. Gjerde
`I
`teaches
`a
`“non-alkylated
`poly(styrene-
`divinylbenzene)” stationary phase ...................................................... 75
`XII. Conclusion .................................................................................................. 76
`
`
`3.
`
`4.
`
`5.
`
`XI.
`
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`
`
`I, David Hornby, Ph.D., hereby declare as follows.
`I.
`
`Introduction
`1.
`
`I am over the age of eighteen (18) and competent to make this
`
`declaration.
`
`2.
`
`I have previously provided a declaration dated September 29, 2017,
`
`(EX1002) in these proceedings in support of MODERNATX, INC.’S (“MODERNA”)
`
`Petition for the above-captioned inter partes review (IPR). The opinions I
`
`expressed in my first Declaration remain unchanged.
`
`3. My professional background and qualifications discussed in ¶¶6-13 of
`
`EX1002, remain unchanged.
`
`4.
`
`I am being compensated for my time in connection with this IPR at
`
`my standard consulting rate, which is $450 per hour.
`
`5.
`
`I understand that this Declaration accompanies the Petitioner’s Reply
`
`to the Patent Owner’s Response dated July 18, 2018.
`
`6.
`
`In preparing this Declaration, I have reviewed the ’340 patent and
`
`each of the documents cited herein, in light of general knowledge in the art. In
`
`formulating my opinions, I have relied upon my experience, education, and
`
`knowledge in the relevant art. In formulating my opinions, I have also considered
`
`the viewpoint of a person of ordinary skill in the art (“POSA”) (i.e., a person of
`
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`ordinary skill in the field of purifying nucleic acids, as defined further below in §
`
`V) prior to December 22, 2006.
`
`II.
`
`Summary of opinions
`7. Claims 1-26 of the ’340 patent are directed to methods of
`
`chromatographic purification of RNA using a porous non-alkylated
`
`polystyrenedivinylbenzene (“PSDVB”) material as the stationary phase. EX1001,
`
`19:57-22:29. In my first declaration (EX1002), I explained that claims 1-26 of the
`
`’340 patent would have been obvious to a POSA, even in light of CureVac’s
`
`asserted evidence regarding objective indicia. EX1002, ¶¶147-288. I also
`
`explained that claims 1-5, 8, 10-22, and 26 are anticipated by Gjerde I. EX1002,
`
`¶¶79-142. My opinions about these grounds have not changed, but I offer the
`
`following additional explanations.
`
`8. A POSA would have had a reason to purify Zhang’s antisense agents,
`
`such as AS-ODNs, ribozymes and siRNA, on a preparative scale using Lloyd’s
`
`method. Zhang specifically provided examples of antisense agents, including
`
`“antisense oligonucleotides,” “ribozymes,” and “small interfering RNAs,” and
`
`taught that large quantities of antisense agents will be needed for “more
`
`preclinical and clinical investigations” in “the near future.” EX1002, ¶¶160, 161;
`
`EX1038, 11:1, 22:4. A POSA would have used Lloyd’s method to purify Zhang’s
`
`antisense RNA on a preparative scale because Lloyd taught that IP RP HPLC
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`method was “an obvious choice” for purifying “oligonucleotides” for “antisense
`
`therapy.” EX1002, ¶162; EX1005, 5:1:2. And a POSA would have known that
`
`RNA was used in “antisense therapy,” and that the term “oligonucleotide” is
`
`generic to both, DNA and RNA. EX1002, ¶158; EX1005, 5:1:2; EX1038,
`
`11:Introduction; EX1064, 90.
`
`9.
`
`Further, a POSA would have understood that Lloyd’s method can
`
`successfully separate a complex mixture of oligonucleotides using a porous
`
`stationary phase. EX1005, 3:1:1, 3:2:1, 4:Figure 1. And Lloyd expressly suggested
`
`using the PLPR-S columns, which had the necessary loading capacity, for
`
`preparative purification. EX1005, 3:2:2, 5:2:1, 6:1:2. Moreover, the existence of
`
`other alternatives of stationary phases in the art would not have made a method
`
`using a porous support non-obvious. And given Lloyd’s teachings and the general
`
`state of the art, a POSA would have reasonably expected that the methods of Lloyd
`
`would have been successful for purifying antisense RNA. EX1002, ¶21; EX1008,
`
`1:2:1-2; Abstract; EX1004, 3:41-52; EX1062, 1:2:2; EX1006, 14:27-30; EX1015,
`
`Abstract; EX1023, 4:1:2; EX 1005, 1:Abstract, 225:2:1; Fig. 1; EX1024, 105:2:1.
`
`10.
`
`In addition, in reaching my opinions regarding obviousness, I
`
`considered whether there were any objective indicia (i.e., objective evidence) of
`
`non-obviousness. I concluded that there is no objective evidence that supports
`
`
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`

`

`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`non-obviousness of the methods claimed in the ’340 patent, as I explain in detail
`
`in Section X.
`
`11. Further, claims 1-5, 8, 10-22, and 26 are anticipated by Gjerde I. The
`
`proper construction of the term “porous reverse phase” is “a reverse phase that is
`
`characterized by pores.” This is a definition that the ’340 patent gave to this term.
`
`EX1001, 7:1-3. Under this claim construction, Gjerde I anticipates claims 1-5, 8,
`
`10-22, and 26 because Gjerde I expressly states that the stationary phase used for
`
`segregation of RNA “can be porous”—i.e., characterized by pores. EX1004, 7:46.
`
`But even under the construction that requires that the pores are of a size such that
`
`RNA can enter the pores, Gjerde I teaches a porous stationary phase. Indeed,
`
`Gjerde I teaches a monolith that contains “a unitary structure having through pores
`
`or interstitial spaces which allow eluting mobile phase and analyte to pass
`
`through.” EX1004, 7:53-64. Thus, Gjerde I expressly teaches that RNA enters the
`
`through-pores of the monolith. Thus, Gjerde I anticipates claims 1-5, 8, 10-22, and
`
`26.
`
`III. List of documents I considered in formulating my opinions
`12.
`In formulating my opinions, I have considered all the references and
`
`documents cited herein, including those listed below.
`
`
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`

`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Exhibit #
`
`Description
`
`1001
`
`1004
`
`1005
`
`1006
`
`1007
`
`1008
`
`1009
`
`1010
`
`1011
`
`1012
`
`1013
`
`Ketterer, T., et al., “Method for Purifying RNA on a Preparative
`Scale By Means of HPLC,” U.S. Patent No. 8,383,340 (filed on
`October 20, 2009; issued on February 26, 2013)
`Gjerde, D., et al., “Method and System for RNA Analysis by
`Matched Ion Polynucleotide Chromatography,” U.S. Patent No.
`6,576,133 (filed on January 2, 2001; issued on June 10, 2003)
`(Gjerde I)
`Lloyd, L, et al., “Rigid polymerics: the future of oligonucleotide
`analysis and Purification,” J. Chromatogr. A 1009: 223-230 (2003)
`Gjerde, D., et al., “Polynucleotide Separations on Polymeric
`Separation Media,” U.S. Patent No. 6,066,258 (filed on October 30,
`1998; issued on May 23, 2000) (Gjerde II)
`Bonn, G., et al., “Nucleic Acid Separation on Alkylated Nonporous
`Polymer Beads,” U.S. Patent No. 5,585,236 (filed on November 17,
`1993; issued on December 17, 1996)
`Azarani, A. and Hecker, K.H., “RNA analysis by ion-pair reversed-
`phase high performance liquid chromatography,” Nucleic Acids Res.
`29: 1-9 (2000)
`Huck, C.W., et al., “Polystyrene/Divinylbenzene Based Monolithic
`and Encapsulated Capillary Columns for the Analysis of Nucleic
`Acids by High-Performance Liquid Chromatography-Electrospray
`Ionization Mass Spectrometry,” Eng. Life Sci. 5: 431-432 (2005)
`Huck, C. W. and Bonn, G.K., “Poly(Styrene-Divinylbenzene) Based
`Media for Liquid Chromatography,” Chem. Eng. Technol. 28: 1457-
`1472 (2005)
`Bidlingmeyer, B. and Wang, Q., “Additives for reversed-phase
`HPLC mobile phases,” U.S. Patent Application Publication No.
`2005/0011836 (filed on July 17, 2003; published on January 20,
`2005)
`Oefner, P.J. and Huber, C.G., “A decade of high-resolution liquid
`chromatography of nucleic acids on styrene–divinylbenzene
`copolymers,” J. Chromatogr. B 782:27-55 (2002)
`Huber, C., et al., “Method And Apparatus For Separating
`Polynucleotides Using Monolithic Capillary Columns,” WO
`2001/055713 (filed on January 25, 2001; published on August 2,
`2001)
`
`- 5 -
`
`
`
`
`
`
`

`

`Exhibit #
`
`1014
`
`1015
`
`1016
`
`1017
`
`1018
`
`1019
`
`1020
`
`1021
`1022
`
`1023
`
`1024
`
`
`
`
`
`
`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Description
`
`Viklund, C., et al., “Monolithic, 'Molded', Porous Materials with
`High Flow Characteristics for Separations, Catalysis, or Solid-Phase
`Chemistry: Control of Porous Properties during
`Polymerization,” Chem. Mater. 8: 744-750 (1996)
`McFarland, G. D. and Borer, P.N., “Separation of oligo-RNA by
`reverse-phase HPLC,” Nucleic Acids Res. 7: 1067-1080 (1979)
`Morgan, R.L. and Celebuski, J. E., “Large-scale purification of
`haptenated oligonucleotides using high-performance liquid
`chromatography,” J. Chromatogr. 536: 85-93 (1991)
`Nielsen, M.D., et al., “High-performance liquid chromatography
`purification of 26-bp serial analysis of gene expression ditags results
`in higher yields, longer concatemers, and substantial time savings,”
`Anal. Biochem. 313: 128-132 (2003)
`Oberacher, H., “Capillary monoliths for the analysis of nucleic
`acids by high-performance liquid chromatography–electrospray
`ionization mass spectrometry,” Trends in Anal. Chem. 21: 166-174
`(2002)
`Necina, R., et al., “Method And Device For Isolating And Purifying
`A Polynucleotide of Interest On A Manufacturing Scale,” WO
`2003/051483 (filed on December 16, 2002; published on June 26,
`2003)
`Oberacher, H., et al., “Characterization of some physical and
`chromatographic properties of monolithic poly(styrene–co-
`divinylbenzene) columns,” J. Chromatogr. A 1030: 201-208 (2004)
`File History for U.S. Patent No. 8,383,340
`Gjerde, D., et al., “Process For Performing Polynucleotide
`Separations,” U.S. Patent No. 6,156,206 (filed on June 2,1999; issued
`on December 5, 2000)
`Georgopoulos, D.E. and Leibowitz, M.J., “Use of high-performance
`liquid chromatographic fractionation of large RNA molecules in the
`assay of group I intron ribozyme activity,” J. Chromatogr. A 868:
`109-114 (2000)
`Chromatography Products from Polymer Laboratories, Catalog Issue
`3, 2004-2005
`
`- 6 -
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`

`

`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Exhibit #
`
`Description
`
`1025
`
`1026
`
`1027
`
`1028
`1029
`
`1030
`
`1031
`
`1032
`
`1033
`
`1034
`
`1035
`
`1036
`
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`Švec, F., “Chapter 2: Organic Polymer Support Materials,” in HPLC
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`Chromatography of Oligonucleotides,” Anal. Biochem. 145: 47-56
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`Putral, L.N., et al., “RNA Interference for the Treatment of Cancer,”
`Drug News Perspect. 19: 317-324 (2006)
`“Chapter 26: An Introduction to Chromatographic Separations,” and
`“Chapter 28: High-Performance Liquid Chromatography,” in
`Principles of Instrumental Analysis, 5th Ed., pp. 674-696, 725-765,
`Skoog, Holler and Nieman (Eds.) Saunders College Publishing 1992
`Taylor, P.D., et al., “MIPC Column Cleaning System and Process,”
`U.S. Patent No. 6,136,195 (filed April 2, 1999; published October
`24, 2000)
`Glossary from Molecular Biology of the Cell, 3rd Ed., pp. G-1 to G-
`23, Alberts et al. (Eds.) Garland Publishing 1994
`Rathore, A.S. and Velayudhan, A., “Chapter 1: An Overview of
`Scale-Up in Preparative Chromatography,” in Scale -Up and
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`pp. 5-36, Marcel Dekker, Inc. 2003
`
`- 7 -
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`

`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Exhibit #
`
`Description
`
`1037
`
`1038
`
`1039
`
`1040
`
`1041
`
`1042
`
`1043
`
`1044
`
`1045
`
`1046
`
`1047
`
`1048
`
`Lloyd, L.L., et al., “Oligonucleotide analysis by anion exchange
`HPLC,” Bioseparation 2: 207-215 (1991)
`Zhang, Y.C., et al., “Antisense Inhibition – Oligonucleotides,
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`(Eds.), pp. 11-34, Humana Press, Inc. 2005
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`RNA,” Nature 418: 252-258 (2002)
`Steitz, T.A. and Moore, P.B., “RNA, the first macromolecular
`catalyst: the ribosome is a ribozyme,” TRENDS in Biochem. Sci. 28:
`411-418 (2003)
`Kieft, J.S. and Batey, R.T., “A general method for rapid and
`nondenaturing purification of RNAs,” RNA 10: 988–995 (2004)
`Green, P., et al., “The Role of Antisense RNA in
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`Scott, W.G. and Klug, A., “Ribozymes: structure and mechanism in
`RNA catalysis,” TIBS 21: 220-224 (1996)
`Hölzl, G., et al., “Analysis of Biological and Synthetic Ribonucleic
`Acids by Liquid Chromatography-Mass Spectrometry Using
`Monolithic Capillary Columns,” Anal. Chem. 77: 673-680 (2005)
`Premstaller, A., et al., “High-Performance Liquid Chromatography-
`Electrospray Ionization Mass Spectrometry of Single- and Double-
`Stranded Nucleic Acids Using Monolithic Capillary Columns,” Anal.
`Chem. 72:4386–4393 (2000)
`Huber, C.G., et al., “Mutation detection by capillary denaturing
`high-performance liquid chromatography using monolithic columns,”
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`Deshmukh, R.R., et al., “Purification of Antisense Oligonucleotides,”
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`http://www.fam.ulusiada.pt/downloads/biblioteca/boletins/listad/mai
`o/internationallaboratorynews1.pdf
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`Exhibit #
`
`1049
`
`1050
`
`1051
`
`1052
`
`1053
`
`1054
`
`1055
`
`1056
`
`1057
`
`1058
`
`
`
`
`
`
`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Description
`
`Pavco, P., et al., “Nucleic Acid-Mediated Treatment of Diseases Or
`Conditions Related To Levels Of Vascular Endothelial Growth
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`2004)
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`of Nov. 13, 2006, available at
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`bs.com/cgi-bin/fillform?file=prodlit.htm
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`03/074654 (filed on February 20, 2003; published on September 12,
`2003)
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`transcription machinery,” Biochem. Soc. Symp. 73: 203-216 (2006)
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`transcriptional modification of tRNA,” Nucleic Acids Res. 26: 1481–
`1486 (1998)
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`Molecular Biology,” “Chapter 5. Replication, Maintenance, and
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`296:1260-1263 (2002)
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`May 31, 1994)
`Transcript from the Deposition of František Švec, Ph.D. (September
`26, 2018)
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Exhibit #
`
`Description
`
`1059
`
`1060
`
`1061
`
`1062
`
`1063
`
`1064
`
`1065
`
`1066
`
`1067
`
`1068
`
`1069
`
`1071
`
`1072
`
`2001
`
`“Isis Pharmaceuticals Initiates Phase II Clinical Trial of ISIS 104838,
`A TNF-Alpha Inhibitor, in Rheumatoid Arthritis,” Isis
`Pharmaceuticals Press Release (March 13, 2002)
`Gniadecki, R., et al., “Trends and Developments in the
`Pharmacological Treatment of Psoriasis,” Acta. Derm. Venereol.
`82:401-410 (2002)
`Kurreck, J., “Antisense technologies Improvement through novel
`chemical modifications,” Eur. J. Biochem. 270:1628-1644 (2003)
`Gelhaus, S., et al., “Rapid purification of RNA secondary
`structures,” Nucleic Acids Research 31:e135 (2003)
`Huber, C.G., et al., “High-resolution liquid chromatography of DNA
`fragments on non-porous poly(styrene-divinylbenzene) particles,”
`Nucleic Acids Research 21:1061-1066 (1993)
`Matthews, C. and van Holde, K., “Chapter 4. Nucleic Acids,” in
`Biochemistry, The Benjamin/Cumming Publishing Company, Inc.
`1996
`Gooding, K. and Regnier, F., “Chapter 2. Organic Polymer Support
`Materials” in HPLC of Biological Macromolecules, Marcel Dekker,
`Inc. 2002
`Swiderski, P., et al., “Polystyrene Reverse-Phase Ion-Pair
`Chromatography of Chimeric Ribozymes,” Analytical Biochemistry
`216:83-88 (1994)
`Transcript from the Deposition of Alexander Schwenger, Ph.D.
`(October 4, 2014) Confidential
`Transcript from the Deposition of Mariola Fotin-Mleczek, Ph.D.
`(October 3, 2018) Confidential
`Transcript from the Deposition of Moritz Thran, Ph.D. (October 2,
`2018) Confidential
`Voet, D. and Voet, G., “Chapter 28. Nucleic Acid Structures and
`Manipulation,” “Chapter 29. Transcription,” and “Chapter 30.
`Translation,” in Biochemistry, John Wiley and Sons, Inc. 1990
`Sambrook, J., Fritsch, E., and Maniatis, T., “Chapter 7. Extraction,
`Purification, and Analysis of Messenger RNA from Eukaryotic
`Cells,” in Molecular Cloning A Laboratory Manual, 2nd Edition,
`Cold Spring Harbor Laboratory Press 1989
`Declaration of František Švec, Ph.D.
`
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Exhibit #
`
`Description
`
`2002
`2003
`
`2004
`
`2005
`
`2006
`
`2007
`
`2008
`
`2009
`
`2010
`
`2012
`2013
`2014
`2016
`2017
`2018
`2019
`2020
`
`2021
`
`2022
`
`2023
`
`Curriculum Vitae of František Švec, Ph.D.
`Martins et al. “Ribonucleic acid purification,” J Chromatogr A.
`2014, 1355, 1-14.
`Snyder, L. et al. “Introduction to Modern Liquid Chromatography,”
`2nd ed. Wiley, Hoboken, NJ, USA, 1979, Chapters 1, 2, 5, 7, 8, 10,
`11, 13, and 14.
`Turkova, J. et al. “Affinity Chromatography,” J. Chromatogr. 1974,
`91, 267-291, Chapters 2 and 6.
`Neue, U.D. HPLC Columns. Theory, Technology, and Practice:
`Chapter 10, Wiley-VCH, New York, 1997.
`Neue, U.D. HPLC Columns. Theory, Technology, and Practice:
`Chapter 2, Wiley-VCH, New York, 1997.
`Neue, U.D. HPLC Columns. Theory, Technology, and Practice:
`Chapter 4, Wiley-VCH, New York, 1997.
`Rodrigues, A.E., et al. “Importance of intraparticle convection in the
`performance of chromatographic processes,” J. Chromatogr., 1992,
`590, 93-100.
`Rodrigues, A.E., et al. “Peak resolution in linear chromatography -
`Effects of intraparticle convection,” J. Chromatogr.,1993, 653, 189-
`198.
`Declaration of Mariola Fotin-Mleczek, Ph.D.
`Declaration of Moritz Thran, Ph.D.
`Declaration of Alexander Schwenger, Ph.D.
`Second Declaration of František Švec, Ph.D., including updated
`Curriculum Vitae of Mariola Fotin-Mleczek, Ph.D.
`Curriculum Vitae of Moritz Thran, Ph.D.
`Curriculum Vitae of Alexander Schwenger, Ph.D.
`Nguyen, T. et al. “High-Resolution Preparative-Scale Purification of
`RNA Using the Prep Cell” Analytical Biochemistry, 1999, 269, 216–
`218
`Phillips, M. et al. Antisense Therapeutics: Chapter 1, Humana Press,
`New Jersey, 2005.
`Ettre, L.S. “Nomenclature for chromatography”, Pure & Appl.
`Chem., 1993, 65, 819-872.
`Ugelstad, U.S. Patent No. 4,563,510, “Process for the preparation of
`polymer latex,” issued Jan. 7, 1986.
`
`- 11 -
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`

`

`Exhibit #
`
`2024
`
`2025
`
`2026
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`2027
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`2028
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`2029
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`2030
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`2031
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`2032
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`2033
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`2034
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`2035
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`
`
`
`
`
`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Description
`
`Waghmare, S. et al. “Studying the mechanism of RNA separations
`using RNA chromatography and its application in the analysis of
`ribosomal RNA and RNA:RNA interactions”, Journal of
`Chromatography A, 2009, 1216, 1377–1382.
`Gjerde, D. et al. RNA Purification and Analysis” Wiley-VCH Verlag
`GmbH & Co., Weinheim, 2009, Chapter 3.
`Dickman, M. “Enrichment and analysis of RNA centered on ion pair
`reverse phase methodology”, RNA, 2006, 12, 691-696.
`Koolman, J. et al. Color Atlas of Biochemistry: Biomolecules,
`second edition, Thieme Verlag, Stuttgart, Germany 2005.
`Li, Y. et al. “Kinetics of RNA Degradation by Specific Base
`Catalysis of Transesterification Involving the 2’-Hydroxyl Group”, J.
`Am. Chem. Soc., 1999, 121, 5364-5372.
`Haupt, W. et al. “Comparison of Several High-Performance Liquid
`Chromatography Techniques for the Separation of
`Oligodeoxynucleotides According to Their Chain Lengths”, Journal
`of Chromatography, 1983, 260, 419-427.
`Horvath, C. et al. “Enhancement of Retention by Ion-Pair Formation
`in Liquid Chromatography with Nonpolar Stationary Phases”,
`Analytical Chemistry, 1977, 49, 2295-2305.
`Kariko, K. et al. “Generating the optimal mRNA for therapy: HPLC
`purification eliminates immune activation and improves translation
`of nucleoside-modified, protein-encoding mRNA”, Nucleic Acids
`Research, 2011, 1-10.
`Anderson, B. et al. “Incorporation of pseudouridine into mRNA
`enhances translation by diminishing PKR activation”, Nucleic Acids
`Research, 2010, 38, 5884-5892.
`Kariko, K. et al. “Incorporation of Pseudouridine Into mRNA Yields
`Superior Nonimmunogenic Vector With Increased Translational
`Capacity and Biological Stability”, Mol Ther., 2008, 16, 1833–1840.
`Nallagatla, S. et al. “RNA structure and regulation of innate
`immunity through protein kinase PKR”, Curr Opin Struct Biol.,
`2011, 21, 119–127.
`Shimabukuro-Vornhagen, A. et al. “Cytokine release syndrome”,
`Journal for ImmunoTherapy of Cancer 2018, 6, 1-14.
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`Exhibit #
`
`Description
`
`2036
`
`2037
`
`2039
`
`2040
`
`2041
`
`Tanaka, T. et al. “IL-6 in Inflammation, Immunity, and Disease”,
`Cold Spring Harb Perspect Biol 2014, 6:a016295, 1-16.
`Guillot, L. et al. “Involvement of Toll-like Receptor 3 in the Immune
`Response of Lung Epithelial Cells to Double-stranded RNA and
`Influenza A Virus”, The Journal of Biological Chemistry, 2005, 280,
`5571–5580.
`Chow, F. et al. “Synthesis of oligodeoxyribonucleotides on silica gel
`support”, Nucleic Acids Research, 1981, 9, 2807-2817.
`Schott, H. et al. “Single-step elongation of oligodeoxynucleotides
`using terminal deoxynucleotidyl transferase”, Eur. J. Biochem. 1984,
`143, 613-620.
`Schott, H. et al. “Column Chromatographic Purification of
`Guanylate-Rich Synthetic Oligodeoxyribonucleotides”, Journal of’
`Chromatography, 1987, 389, 165-176.
`
`IV. State of the art before December 22, 2006
`13.
`In addition to my comments on the state of the art that I provided in
`
`¶¶30-64 of my first declaration (EX1002), I provide the following additional
`
`comments.
`
`A. Overview of RNA
`14. Ribonucleic acid (RNA) is a polymeric molecule essential for life.
`
`EX1071, 791:2:4-5, 792:1:1-4. Like its deoxyribonucleic acid counterpart (DNA),
`
`RNA is a polymeric chain of nucleoside units connected via a negatively-charged
`
`phosphodiester backbone. EX1071, 847:2:1; EX1058, 39:7-20, 40:5-6. Like DNA,
`
`RNA contains four purine and pyrimidine bases: adenine (A), guanine (G),
`
`cytosine (C), and uracil (U) (DNA contains thymine (T) instead of U). EX1071,
`
`- 13 -
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`IPR2017-02194
`Second Declaration of David Hornby, Ph.D (EX1070)
`
`792:2:Figure 28-1; EX1058, 39:21-40:1. The primary difference between RNA
`
`and DNA is in the structure of the nucleoside unit: RNA contains a ribose unit,
`
`while DNA has a deoxyribose unit. EX1071, 847:2:1; EX2016, ¶10.
`
`15. The functional world of RNA is very diverse. Some RNA molecules
`
`play an active role within cells by modifying other RNAs and catalyzing
`
`biological reactions; these include small nuclear RNAs (snRNA) and ribozymes.
`
`EX1071, 883:2:2. Others, such as RNAi, long non-coding RNAs, and enhancer
`
`RNAs, control gene expression. EX1055, 130:4 and 246:2; EX1056, 1260:1. RNA
`
`is also involved in protein synthesis, where messenger RNA (mRNA) molecules
`
`direct the assembly of proteins on ribosomes. EX1071, 853:2:3. This process uses
`
`mRNA to convey genetic information from DNA that specifies the amino acid
`
`sequence of proteins, transfer RNA (tRNA) to deliver amino acids to the
`
`ribosome, and ribosomal RNA (rRNA) to link amino acids together to form
`
`proteins. EX1055, 231:1. And both Dr. Fotin-Mleczek and Dr. Švec admitted that
`
`there are many types of RNAs, and they include, for example, “messenger RNA,”
`
`“[l]ong chain noncoding RNA, Circular RNA, MicroRNA, SiRNA, … tRNA,
`
`ribosomal RNA”. EX1068, 33:3-8; EX1058, 27:2-30:18.
`
`16. RNA molecules also vary in length considerably. Some RNAs are
`
`shorter than 200 nucleotides (nt) in length, and include, e.g., tRNAs, ribozymes,
`
`some rRNAs, and small interfering RNAs (siRNA). EX1038, 18:4; EX1071,
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`IPR2017-02194
`Second Declaration of Dav