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: [email protected]
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