The future of liquid biopsy is here

The future of liquid biopsy is here

CTC Ion Torrent LiquidBiopsy Platform p4 DNA cfdna MagMAX Cell-Free DNA Isolation Kit p6 MagMAX Sample Preparation p6 KingFisher p7 Total Exosome Isolation p7 Ion S5 / Ion S5 XL p8 Ion PGM p9 Ion AmpliSeq p10 cfdna Oncomine cfdna Assay p12 p14 p15 PCR TaqMan Mutation Detection Assays p16 PCR QuantStudio 3D PCR p17 NanoDrop One & Qubit 3 Fluorometer p18 Thermo Fisher Cloud p19

liquid biopsy biopsy Circulating Tumor Cell: CTC DNA cell-freedna: cfdna CTC 10 ml cfdna DNA cfdna CTC cfdna CTC 回収 CTC 染色 計数 10ml 全血 LiquidBiopsy Flow Chip EVOS FL Cell Imaging System がん関連遺伝子領域増幅 NGS ライブラリ作製 変異解析 (NGS) P4-5 LiquidBiopsy Blood Collection Kit P4-5 cfdna 抽出 cfdna 自動抽出 Ion AmpliSeq Panel Oncomine cfdna Assays P10-13 Ion Torrent System P8-9 14-15 変異定量 ( デジタル PCR) MagMAX cfdna Isolation Kits P6 KingFisher Flex P7 QuantStudio 3D P16-17

CTC Ion Torrent LiquidBiopsy Platform Ion Torrent LiquidBiopsy Platform CTC CTC CTC EpCAM EMT 7.5-10 ml CTC cfdna Germline Control White Blood Cell 96 CTC Ion Torrent LiquidBiopsy Instrument Graphical User Interface 82 cm Slide/Chip Tray Assembly 4 Pumps 82 cm 58 cm CTC CTC EDTA2K 7.5-10mL cfdna 4

1,000 80-100% 3-5 cells 22 20 cells/ml CTC CTC cells/ml 100 10 1 H H H H H H HH H H H 0 1 10 100 1,000 cells/ml 7.5mL MCF-73, 9, 30, 90, 300, 900 cells/ml LiquidBiopsy Platform 1 target cell/ml 1000 cells/ml Iohexol PBS Variants Variant frequency Position Gene Reference Variant WBCs CTCs cfdna chr3:178917005 PIK3CA A G 0.496 0.5 0.6378 chr4:1806099 FGFR3 A G Not present 0.0119 0.0156 chr4:1807894 FGFR3 G A 0.996 1 0.9969 chr4:55141055 PDGFRA A G 1 1 0.994 chr4:55152040 PDGFRA C T 0.996 0.9819 0.996 chr4:55980239 KDR C T 0.339 0.36 0.339 chr5:112175770 APC G A 1 1 1 chr5:149433596 CSF1R TG GA 0.988 0.995 0.988 chr7:55249063 EGFR G A 0.5089 NC 0.517 chr11:534242 HRAS A G 0.507 0.498 0.5308 chr13:28610183 FLT3 A G 1 1 1 chr18:48586344 SMAD4 C T 0.506 0.469 0.468 CTC cfdna DNA LiquidBiopsy PlatformCTC, cfdna WBCs) 3DNA Ion AmpliSeq Cancer Hotspot Panel v2 Ion 318 Chip 1 COSMIC COSM760 WBC LiquidBiopsy Instrument 2 1 CVLB1-001 19,500,000 LiquidBiopsy Blood Collection Kit 1 A28171 9,400 CTC LiquidBiopsy Reagents and Consumables Kit, Part 1 8 A28186 142,800 LiquidBiopsy Reagents and Consumables Kit, Part 2 8 A32500 282,000 EpCAM Capture Antibody 8 A32596 9,000 Her2 Capture Antibody 8 A32597 9,000 Trop2 Capture Antibody 8 A32598 9,000 Bi Label Epcam + Trop2 Capture Antibodies 8 A32599 9,000 Bi Label Epcam + Her2 Capture Antibodies 8 A32600 9,000 Bi Label Her2 + Trop2 Capture Antibodies 8 A32601 9,000 Tri Label Epcam + Her2 + Trop2 Capture Antibodies 8 A32602 9,000 5

DNA cfdna MagMAX Cell-Free DNA Isolation Kit Applied Biosystems MagMAX Cell-Free DNA Isolation Kit cfdna cfdna 500 µl-10 ml 15-50 µl KingFisher7 40 K DNA 170 bp cfdna DNA cfdna 4 ml 50 µl 50 bp DNA 2 ng/µl 60 µl Dynabeads MyOne Silane DNA / 50 µl 1 µl1 µl DNA Agilent Bioanalyzer 2100 HS dsdna 100-750 bp DNA 100% MagMAX Sample Preparation MagMAX Sample Preparation RNA RNA RNA 20 µl RNasePCR KingFisher RNA small RNA RNA mirna RNA DNA DNA/RNA / MagMAX for Stabilized Blood, Tempus MagMAX for Stabilized Blood, Paxgene MagMAX Blood RNA Isolation MagMAX mirvana Total RNA Isolation Kit MagMAX DNA Multi-Sample Ultra Kit MagMAX Cell-Free DNA Isolation Kit MagMAX Pathogen RNA/DNA Kit MagMAX Cell-Free DNA Isolation Kit 1 A29319 80,000 MagMAX for Stabilized Blood Tubes RNA Isolation Kit, 96 4451893 103,100 compatible with Tempus Blood RNA Tubes MagMAX for Stabilized Blood Tubes RNA Isolation Kit, 96 4451894 102,300 compatible with PAXgene Blood RNA Tubes MagMAX-96 Blood RNA Isolation Kit 96 AM1837 75,600 MagMAX mirvana Total RNA Isolation Kit 96 A27828 65,000 MagMAX DNA Multi-Sample Ultra Kit 500 A25597 254,700 MagMAX Pathogen RNA/DNA Kit 480 4462359 175,000 6

純度 累積率 純度 累積率 KingFisher Thermo Scientific KingFisher MagMAX Sample Preparation 96 MagMAX Preperlation Kit BindIt KingFisher Duo Prime 30-5,000 µl 6 12 24 24 96 well KingFisher Flex 20-5,000 µl 24 96 24 96 well PCR KingFisher Duo Prime 5400110 3,000,000 KingFisher Flex 96DW * 5400630 7,040,000 * 3 Total Exosome Isolation Invitrogen Total Exosome Isolation Kit Invitrogen Total Exosome RNA & Protein Isolation Kit RNA [Total Exosome Isolation 試薬 ] [ 超遠心法 ] 100% 90% 80% 70% 60% 50% 40% 100% 90% 80% 70% 60% 50% 40% HeLa Nanosight LM10 300nm 50-150nm 30% 30% 20% 20% 10% 10% 0 0 100 200 300 400 500 600 700 800 粒子サイズ 900 0 0 100 200 300 400 500 600 700 800 粒子サイズ 900 Total Exosome Isolation Reagent 6 ml 4478360 41,500 Total Exosome Isolation Kit 6 ml 4484450 43,700 Total Exosome Isolation Reagent 50 ml 4484452 29,500 Total Exosome Isolation Reagent 6 ml 4484453 43,700 Total Exosome RNA & Protein Isolation Kit 40 4478545 41,500 7

Ion S5 / Ion S5 XL FFPE Ion S5 /S5 XL Ion Torrent DNA RNA Ion AmpliSeq Panel Ion Torrent Oncomine Cancer Assay CNV 400 bp 6 8 Gb 15 200 bp 2.5 400 bp 4 Ion S5 Ion S5 XL Ion 520 Ion 530 Ion 540 Ion 520 Ion 530 Ion 540 300 500 1,500 2,000 6,000 8,000 300 500 1,500 2,000 6,000 8,000 * 200 bp 0.6 1 Gb 3 4 Gb 10 15 Gb 0.6 1 Gb 3 4 Gb 10 15 Gb 400 bp 1.2 2 Gb 6 8 Gb 1.2 2 Gb 6 8 Gb 200 bp 2.5 2.5 2.5 2.5 2.5 2.5 400 bp 4 4 4 4 200 bp 5 8 16.5 1 2.5 5 400 bp 8 17.5 2 4 * >99%/ Aligned BAM Ion S5 Ion S5 Ion OneTouch 2.0 Duo Ion S5 Ion S5 Ion Chef Ion S5 XL Ion S5 XL Torrent Ion OneTouch 2.0 Duo Ion S5 XL Ion S5 XL Torrent Ion Chef 1 S5-001OT2S2 2 13,300,000 1 S5-001CS2 2 17,700,000 1 S5XL-001OT2S2 2 26,100,000 1 S5XL-001CS2 2 29,800,000 8

Ion PGM Ion PGM 3 Ion 314 /316 /318 200 400 bp30 Mb 2 Gb 1 5 Ion 314 v2 Ion 316 v2 Ion 318 v2 130 630 1200 200 bp 30-50 Mb 300-600 Mb 600 Mb-1 Gb 400 bp 60-100 Mb 600 Mb-1 Gb 1.2-2 Gb 40-60 200-300 400-550 200 bp 2.3 3.0 4.4 400 bp 3.7 4.9 7.3 Ion PGM Ion PGM Ion Torrent Ion OneTouch 2.0 Duo 1 PGM11-020S2 2 9,600,000 Ion PGM Ion PGM Ion Torrent Ion Chef 1 PGM11-C01S2 2 13,400,000 9

Ion AmpliSeq Panel Ion AmpliSeq Panel PCR Ion Torrent DNA Ion AmpliSeq Cancer Hotspot Panel v2 2071 NCI 50 2,790 1 ng 1 PCR Ion 314 / 316 / 318 FFPE 50 111-187 bp 154 bp 1 DNA 10 ng 10 ng x 1 10 * >95% ** >90% NA * 20% **1 KRAS BRAF EGFR TP53 PIK3CA CSF1R JAK2 NRAS PTPN11 ERBB2 SRC FGFR3 NPM1 CDKN2A RET HNF1A SMAD4 GNAS PDGFRA MPL ABL1 PTEN FLT3 STK11 SMARCB1 KIT MET NOTCH1 FGFR2 RB1 JAK3 VHL KDR SMO EZH2 HRAS AKT1 ALK MLH1 FBXW7 FRBB4 GNA11 ATM CDH1 IDH1 CTNNB1 APC FGFR1 GNAQ IDH2 Ion AmpliSeq Comprehensive Cancer Panel 409 16,000 4 DNA CDS DNA 1 4 ng DNA 4 PCR 41 Ion 318 FFPE 409 125-175 bp 155 bp 4 DNA 40 ng 10 ng x 4 16 * >90% ** >95% NA * 20% **1 10

Ion AmpliSeq Community Panel RNA web www.thermofisher.com/ampliseq.com 1 ng DNA 500 FFPE CNV TP53 Panel TP53 1 24 BRCA1 and BRCA2 Panel BRCA1, BRCA2 2 167 Colon & Lung Cancer Research Panel v2 NRAS, PTEN, PIK3CA 22 92 AML Research Panel DNMT3A, RUNX1 19 264 RNA Fusion Lung Cancer Research Panel ALK, RET, ROS1,NTRK 70 85 Ion AmpliSeq Custom Panel Ion AmpliSeq Cancer Hotsopt Panel v2 Comprehensive Cancer Panel Inherited Disease Panel Ion AmpliSeq Designer 24-96 97-6,144 6,144 10kb Ion AmpliSeq 3-4 24-96 97-6,144 100 or 200bp 1-3 750 3,000 384well 1,500 6,000 Ion AmpliSeq Cancer Hotspot Panel v2 1 8 4475346 28,000 Ion AmpliSeq Comprehensive Cancer Panel 4 8 4477685 108,000 Ion AmpliSeq Colon and Lung Cancer Research Panel v2 1 750 69,000 Ion AmpliSeq TP53 Panel 2 750 36,000 Ion AmpliSeq BRCA1 and BRCA2 Panel 3 3,000 125,250 Ion AmpliSeq AML Research Panel 4 750 198,000 Ion AmpliSeq RNA Cancer Panel 24 4482572 74,000 Ion AmpliSeq RNA Lung Fusion Cancer Research Panel 8 CP1001 32,000 Ion AmpliSeq Custom Panel 750 3,000 1 750 11

cfdna Oncomine Cell-Free DNA Assay Ion Torrent Oncomine cfdna Assay Ion Torrent Oncomine Knowledgebase Ion Torrent SNVs PCR cfdnacfdna 0.1% 1-20 ng cfdnadna 2 cfdna NSCLC cfdna DNA Oncomine cfdna Assays cfdna Limit of Detection 1 ng 0.6 % 5 ng 0.25 % 10 ng 0.15 % 20 ng 0.1 % 30 ng 0.05 % Oncomine cfdna Assays cfdna 20ng 0.1% cfdna cfdnamagmax Cell-Free DNA Isolation Kit 6 KingFisher 7 12

Oncomine Lung cfdna Assay NSCLC11 169 35 11 ALK,BRAF,EGFR,ERBB2,KRAS, MAP2K1,MET,NRAS,PIK3CA, ROS1,TP53 EGFR T790M, C797S, L858R, Exon 19 del KRAS G12X, G13X, Q61X BRAF V600E ALK Exon 21-25 PIK3CA E545K, H1047R, E542K Oncomine Colon cfdna Assay 14 24449 14 AKT1, BRAF, CTNNB1, EGFR, ERBB2, FBXW7, GNAS, KRAS, MAP2 K1, NRAS, PIK3CA, SMAD4, TP53, APC KRAS/NRAS: G12/G13/Q61 BRAF: V600E PIK3CA: E545K, H1047R TP53: R175H R273H/C/L Recurrent deleterious APC mutations including p.r876*, p.r1114*, p.q1378*, and p.r1450* SMAD4: R361C/H CTNNB1: S45F, T41A Oncomine Breast cfdna Assay 10 15926 10 AKT1, EGFR, ERBB2, ERBB3, ESR1, FBXW7, KRAS, PIK3CA, SF3B1, TP53 PIK3CA: E545K and H1047R AKT1: E17K ESR1: Mutations associated with anti-estrogen resistance TP53: Mutations associated with loss of function ERBB2: Mutations associated with sensitivity to anti-erbb2 therapies Oncomine Lung cfdna Assay 8 A31149 280,000 Oncomine Colon cfdna Assay 8 A31182 220,000 Oncomine Breast cfdna Assay 8 A31183 344,000 Tag Sequencing BC Set 1-24 24BC 10 A31830 160,000 Tag Sequencing BC Set 25-48 24BC 10 A31847 160,000 MagMAX Cell-Free DNA Isolation Kit 1 A29319 80,000 KingFisher Duo Prime 5400110 3,000,000 KingFisher Flex 96DW 5400630 7,040,000 13

96 Ion Xpress Barcode Adaptor Kit Ion Xpress Barcode Adaptor Kit Ion Torrent DNA 96 1 96 1 Ion Xpress Barcode Adapters 1-96 Kit 96BC 10 4474517 980,000 Ion Xpress Barcode Adapters 1-16 Kit 16BC 10 4471250 214,000 Ion Xpress Barcode Adapters 17-32 Kit 16BC 10 4474009 214,000 Ion Xpress Barcode Adapters 33-48 Kit 16BC 10 4474518 214,000 Ion Xpress Barcode Adapters 49-64 Kit 16BC 10 4474519 214,000 Ion Xpress Barcode Adapters 65-80 Kit 16BC 10 4474520 214,000 Ion Xpress Barcode Adapters 81-96 Kit 16BC 10 4474521 214,000 Tag Sequencing BC Set 25-48 24BC 10 A31847 160,000 384 IonCode Barcode Adapters Kit IonCode Barcode Adapters 1-384 Kit 384BC 10 A29751 1,152,000 Ion AmpliSeq Ion AmpliSeq Library Kit 2.0 Ion AmpliSeq Library Kit 2.0 Ion AmpliSeq AmpliSeq Ion AmpliSeq 11 Ion AmpliSeq 96LV 384LV 96 1 96 Ion AmpliSeq Library Kit 2.0 8 4475345 108,000 Ion AmpliSeq Library Kit 2.0 96LV 96 4480441 1,020,000 Ion AmpliSeq Library Kit 2.0 384LV 384 4480442 2,720,000 3 Ion Library Equalizer Kit Ion Library Equalizer Kit 3 PCR Ion Torrent DNA Ion AmpliSeq Library Kit 2.0 Ion Plus Fragment Library Kit Ion Xpress Plus Fragment Library Kit Ion Library Equalizer Kit 96 4482298 68,000 14

Ion Reporter Software Ion Torrent Torrent Suite Software Ion Reporter Software Ion Torrent 3 CNV Ion Reporter Software https://ionreporter.iontorrent.com ID Sign In Launch analysis Next Next Next Launch Analysis 15

PCR TaqMan Mutation Detection Assays Applied Biosystems TaqMan Mutation Detection Assays Competitive Allele-Specific TaqMan PCR castpcr MGB 0.1 1 7 log 3 1 TaqMan Mutation Detection Assay Reference Assay Mutant Assay Mutant Allele Assay EGFR_exlqdels_mu 19 MGB EGFR exon19 deletion Wild type Assay Mutant Assay EGFR Reference Assay EGFR Exon19 Deletion Assay Wild type Allele Assay Gene Reference Assay MGB Mutation Detector TaqMan Mutation Detection Assay ASP: LST: TaqMan LSP: MGB: ASB: MGB FP: RP: * TaqMan Mutation Detection Assays 46 778 150 µl /75 4465804 90,800 TaqMan EGFR Exon 19 Deletions Assay 1 EGFR 19 150 µl/75 4465805 90,800 TaqMan Mutation Detection Reference Assays 46 150 µl /75 4465807 56,200 TaqMan Mutation Detection IPC Reagent Kit 1 kit/ 500 4467538 31,800 *20µL www.thermofi sher.com/taqman TaqMan Mutation Detection Assays 16

PCR QuantStudio 3D PCR PCR PCR Applied Biosystems QuantStudio 3D PCR0.1% 0.1% Applied Biosystems Custom TaqMan SNP Genotyping Assays 20,000 PCR 0.1% Custom TaqMan SNP Genotyping Assays KRAS G12V A. DNA 10% PCR 10.384% 8.235% - 13.076% B. DNA 1% PCR 1.393% 0.772% - 2.507% C. DNA 0.1% PCR 0.142% 0.064% - 0.310% D. QuantStudio 3D PCR EGFR, KRAS, BRAF, PIK3CA, JAK2 www.thermofisher.com/jp-dpcr-assays QuantStudio 3D PCR PCR QuantStudio 3D PCR ProFlex PCR QuantStudio PCR ProFlex PCR QuantStudio 3D ChipLoader 1 QS3D-PF 2 6,400,000 QuantStudio 3D PCR Chips v2 12 A26316 11,880 QuantStudio 3D PCR Master Mix v2 1.5 ml A26358 17,000 5.0 ml A26359 48,000 Custom TaqMan SNP Genotyping Assays Custom TaqMan SNP Genotyping Assays for Rare Mutation Analysis 40x, Medium scale 450 4332077 65,900 17

NanoDrop One & Qubit 3 Fluorometer DNA Thermo Scientific NanoDrop One Invitrogen Qubit 3 Fluorometer NanoDrop One NanoDrop One µl UV 1-2 µl 3 Qubit 3 Fluorometer Qubit 3 Fluorometer 2 DNA RNA Qubit 3 NanoDrop One NO DNA A260 /A280 NO RNA, dntps, NO DNA NO DNA NO 100 ng/µl 1,000 ng/µl YES YES YES YES YES NanoDrop One Qubit 3 Qubit 3 NanoDrop One NanoDrop One NanoDrop One 1 ND-ONE-W 1,750,000 NanoDrop One C 1 ND-ONEC-W 2,050,000 Qubit 3.0 Fluorometer 1 Q33216 201,000 Qubit 3.0 Quantitation Starter Kit 1 Q33217 221,100 Qubit 3.0 NGS Starter Kit 1 Q33218 216,100 18

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For Research Use only. Not for use in diagnostic procedures. 2016 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. BIOANALYZER is a trademark of Agilent Technologies, Inc.. TaqMan is a registered trademark of Roche Molecular Systems Inc., used under permission and license. 2016 12 www.thermofisher.com/jp-tc ION148-A1612OB 108-0023 4-2-8 0120-477-392 jptech@thermofisher.com TEL 03-6832-6980 FAX 03-6832-9584 TEL 03-6832-9300 FAX 03-6832-9580 facebook.com/thermofisherjapan www.thermofisher.com @ThermoFisherJP