WuXi AppTecClinical Virology Platforms and Services

www.wuxiapptec.com http://rsd.wuxiapptec.com

Research Service Division (RSD)Enabling Unbounded Possibilities

Other viruses

Advanced therapies

Genotyping and biomarkers

HBV

Clinical Virology

Genotyping

n Viral load determination, genotyping and sequencing: HBV, HCV and RSV

n Line probe assay: LiPAn Genotyping: pharmacogenomics, IL28B

genotyping, MET exon 14 skipping, etc.n Gene expression: ISGs, fresh tissues,

blood, etc.

Clinical Biomarkers

Clinical Experiences

n HBV • HBsAg, HBsAb, HBeAg, HBeAb and

HBcAb• Serum HBcrAg

n Cytokines and chemokines (ELISA, MSD, Luminex)

n IFNs and ISGs

LumiPulse G1200 (Fujirebio)Electrochemiluminescence

Cobas e 601AID ELISpot Reader

MESO SECTOR S600SpectraMax M2eBio-Plex 200 system (Bio-Rad)

COBAS® AmpliPrepInstrument

3730xl Genetic Analyzer and NextSeq 550QX200 Droplet Digital PCR System

HBV clinical support• DNA VL• Phenotyping• Whole genome sequencing• Sanger sequencing of pol-RT gene• INNO-LiPA HBV Multi-DR assay• HBcrAg• HBV RNA quantification • HBV RNA sequencing• ISGs and cytokines • Human gene testing

HCV clinical support• Deep sequencing (1a, 1b, 2a, 3a, 6a)• Phenotyping• Sanger sequencing• Clonal sequencing• Genotyping• RNA VL

RSV clinical support • Virus identification from NP swabs • Genotyping: qPCR and sequencing• Sanger population and clonal sequencing:

F and G genes• RNA VL: qPCR and plaque assay• Virus isolation: propagation TCID50

titration

Cell therapy support• VCN• RCL• Cytokines• IFN-gamma ELISpot• FACS• ADA• Nab

Other clinical support• NASH• Oncolytic virus• Human genes: CYP2D6, IL28B,

TLR,UGT1A9, UGT1A1, etc. • Gene expression

Services of CAP-certificated Molecular Testing Lab

Biomarkers for NASH, CYP2D6, IL28B, UGT1A9 genotyping,Gene expression by RT-PCR

Cell therapy (VCN, RCL, cytokine), gene therapy, oncolytic viruses

HCV viral load and sequencing, RSV viral load and sequencing, other virus sequencing, NGS, etc.

HBV DNA, HBV pol-RT/core/genome sequencing, HBV RNA quantification, HBV RNA sequencing, HBcrAg, ELISpot, NGS, etc.

QiasymphonyDNA/RNA extraction

Agilent 2100/QIAxcelDNA/RNA QC

ABI 7900HT, QuantStudio™ 6/7Real-Time PCR System

The CAP Accredited Molecular Testing Laboratory

Clinical Virology

HBV Genotyping Assays

Test Method Standard Status

HBV DNA VLDCobas CAP In service

ddPCR CAP Validated

HBV whole genome sequencingSanger CAP

In serviceNGS Non-CAP

HBV pol-RT sequencing Sanger CAP In service

HBV core sequencing Sanger CAP Validated

HBV RNA quantificationqPCR CAP In service

ddPCR CAP Validation on going

HBV RNA sequencing Sanger Non-CAP In service

Test Method Quantitative/qualitative Standard/Status

HBsAg ECL Quantitative

CAP/In service

Anti-HBs ECL Qualitative

HBeAg

ELISA Quantitative

ECL Qualitative,semi-quantitative

Anti-HBe ECL Qualitative,semi-quantitative

Anti-HBc ECL Qualitative,semi-quantitative

Serum HBcrAg LumiPulse G1200 Quantitative

HBV Serology Assays

Other Markers

Test Method Standard Status

HBV phenotypingHBV full-length,

pol and capsid shuttle vectors Non-CAP In service

IFN-gamma ELISpot Non-CAP In service

Immunological markers FACS Non-CAP In service

Cytokines Luminex/MSD/ELISA CAP/Non-CAP In service

ISG gene expression RT-qPCR CAP In service

WuXi HBV Clinical Virology Platform

Clinical Virology

Contact

HBV Phenotyping Assays

Zhengxian Gu Executive Director Mobile: +86 189-3083-0132Tel: 86-21-5046-3753 Email: gu_zhengxian@wuxiapptec.com

n Full-length, pol/RT and core gene replication-competent shuttle vectorsn Fitness test for HBV clinical constructs: >400 constructs evaluatedn Drug susceptibility: >1000 compounds/constructs evaluated

HBV titer (log10copies/ml)

PCR success rate

8-9 100% (2/2)

7-8 100% (3/3)

6-7 100% (3/3)

5-6 75% (3/4)

4-5 100% (4/4)

3-4 100% (4/4)

2-3 33.3% (1/3)

Total 87% (20/23)

GLS4

Sample

-1_B

Sample

-2_C

Sample

-3_B

Sample

-4_B

Sample

-5_C

Sample

-6_C

Sample

-7_B

Sample

-8_B

Sample

-9_C

Sample

-10_C

Sample

-11_B

Sample

-12_B

Sample

-13_C

Sample

-14_C

Sample

-15_B

Sample

-16_B

Sample

-17_C

Sample

-18_C

Sample

-19_C

Sample

-20_B

Sample

-21_D

Sample

-22_A

Sample

-23_A

Sample

-24_A

0

20

40

60

80

100

1201st exp2nd exp3rd exp

EC50

(nM

)

ETV

Sample-1_

B

Sample-2_

C

Sample-3_

B

Sample-4_

B

Sample-5_

C

Sample-6_

C

Sample-7_

B

Sample-8_

B

Sample-9_

C

Sample-10

_C

Sample-11

_B

Sample-12

_B

Sample-13

_C

Sample-14

_C

Sample-15

_B

Sample-16

_B

Sample-17

_C

Sample-18

_C

Sample-19

_C

Sample-20

_B

Sample-21

_D

Sample-22

_A

Sample-23

_A

Sample-24

_A0.00.51.01.52.02.53.03.54.0

1st exp2nd exp3rd exp

EC50

(nM)

Assay reproducibility Minor species sensitivity

Immunological Assays

n Isolation of PBMCn T and B cell ELISpotn FACS assaysn Intracellular staining (ICS)

Other Assays

Oncolytic virusn Immunogenicity tests• NAB detection

n Biodistribution/PK tests• Q-PCR detection with serum, saliva or urine• Plaque assays/TCID50 for detection of infectious virus

shedding• Virus replication in tumor: IHC

n Immune response• Transgene expression/functional assay• Cytokines and inflammation• Immune cell activation

CAR-T/TCR-T• RCL/RCR Assay • Vector copy number • TCR sequencing• T cell phenotype or differentiation • Cytokine or chemokine expression and secretion• LDH (lactate dehydrogenase) assay• IFN-gamma ELISpot • FACS

GLS4

ETV

Ex-vivo stimulation of lymphocytes

ELISpot FACS analysis