1. Monogram Biosciences Pharma Collaborations
A LabCorp Center of Excellence –

2. Monogram Biosciences
Founded as ViroLogic, Nov 1995, South San Francisco, CA
Acquired by LabCorp as wholly-owned subsidiary, Aug 2009
~225 employees
Virology/Oncology
Commercial Assays
Clinical management
Drug/vaccine development
Clinical Reference Laboratory
CLIA/CAP accredited
Science directed
Over 175 peer-reviewed publications
Ranked highly for intellectual property portfolio
Has conducted over 200 projects involving every commercially-available HIV antiretroviral

3. Monogram Biosciences A LabCorp Center of Excellence
LabCorp’s Platform
220,000 customers ,000 employees
1,500 patient centers ,000 samples per day
40 years
Our mission is to be a leading provider of high science laboratory services that support clinical decision-making, enhance the management of laboratory information, and improve patient care.

4. Monogram’s Technologies are Offered Directly at Monogram and through Esoterix Clinical Trials Services, a Division of LabCorp
Bioanalytical Labs
Biomarker Discovery
Companion Diagnostics
Central Labs
Esoteric Testing
Small and large molecule
Method development and validation
LC/MS/MS
Non-GLP discovery services
Cell-based assays
Dedicated labs for clinical trials
Cranford, NJ
Mechelen, Belgium
Beijing, China
Singapore
Broad, validated test menu
Test method harmonization
8 Centers of Excellence
Coagulation
Endocrinology
Oncology
Infectious disease
Anatomic pathology
Molecular pathology
Flow cytometry
Discovery and development
GLP biomarker services
MS-based assays
Global expertise:
Cranford, NJ
Mechelen, Belgium
San Diego, CA
West Trenton, NJ
Discovery, development, commercialization
FDA experience
IVD Trial Participation
Successful PMA submission
40,000 ft2 biorepository
Commitment to Quality
Integrated Data Management
Dedicated Project Management

5. Testing Services Across all Phases of Development
Screening drug candidates
Characterizing resistance
Screening patients for enrollment
Optimizing background therapy
Characterizing treatment failure
Defining clinical cut-offs
Enhancing market positioning
Discovery
Preclinical
Clinical
Post-Marketing

6. HCV Testing Services

7. Monogram HCV Assays and Services
Monogram is designed to be a ‘one stop shop’ to meet virology testing needs for clinical trials
Monogram is applying its legacy of providing accurate, high throughput HIV testing to HCV
Current HCV assays
NS3: Sequencing & Phenotyping
NS5b: Sequencing & Phenotyping
NS5A: Sequencing & Phenotyping
IL28B SNP Genotyping
Subtyping/Genotyping and Viral Load
Deep sequencing and clonal analysis

8. GenoSure® HCV – NS3/4a sequencing
Monogram’s GenoSure HCV assay is now available for use in the clinic
Designed to help determine patient susceptibility to the commercially available HCV drugs
Validated to measure VL>2000 copies/ml

9. Monogram HIV Assays and Services for Clinical Research and Development
Protease/Reverse Transcriptase Inhibitors
PhenoSense® (phenotype)
GenoSure® MG (genotype)
PhenoSense® GT≤ (combination phenotype/genotype)
Entry Inhibitor Susceptibility (T20)
GenoSure PRIme® – genotyping PR/RT and Integrase in a single assay
Trofile®, Trofile® DNA (Co-receptor Tropism)
Integrase Susceptibility (phenotype; genotype)
Assembly Inhibitor Susceptibility (phenotype; genotype)
Replication Capacity & Sequencing Subtype
Neutralizing Antibodies – for HIV and Influenza
Clonal Analyses
PhenoScreen® Novel Drug Testing
Database queries
Viral Load (Roche COBAS/Ampliprep TaqMan 2.0, Abbott Real Time)

10. HIV PR/RT Resistance Testing

11. PhenoSense HIV Cell Assay
Resistance Test Vector
DNA
A-MLV env
DNA +
Transfection
Infection
293-A
Pseudovirions
PR inhibitors added
RT inhibitors added

12. Patient virus exhibits reduced susceptibility to the drug
PhenoSense HIV Data
Patient virus exhibits reduced susceptibility to the drug
Patient:
Control:

13. GenoSure MG Report
Monogram’s GenoSure MG report combines LabCorp’s genotyping capabilities (for faster turnaround), and Monogram’s proprietary algorithm…

14. Measure of Drug Susceptibility Prediction of Drug Susceptibility
Parallel Phenotype/Genotype Testing PhenoSense GT Assay (PSGT)
Patient virus
Phenotype
Genotype
RT-PCR
PR-RT DNA
Vector Assembly
Resistance Test Vector
Sequencing
Transfection
Recombinant Virus
Resistance Mutations
Infection
Interpretation
Measure of Drug Susceptibility
Prediction of Drug Susceptibility
Bioinformatics

15. Drug Resistance Testing: Genotype vs Phenotype
Benefits and Limitations
Genotyping
Generates viral sequence (population)
Identifies virus mixtures
mix of susceptible and resistant virus species
example: K103K/N
Major Limitation: Interpretation of complex mutational patterns or reliance on algorithm
Immature algorithms for new agents 15

16. Drug Resistance Testing: Genotype vs Phenotype
Benefits and Limitations
Genotyping
Phenotyping
Generates viral sequence (population)
Identifies virus mixtures
mix of susceptible and resistant virus species
example: K103K/N
Major Limitation: Interpretation of complex mutational patterns or reliance on algorithm
Immature algorithms for new agents
Direct measure of susceptibility
Ease of interpretation
Demonstrates effect of complex combinations of mutations
Resistance and Cross-Resistance
Re-sensitization
Partial Activity
Replication Capacity 16

17. Parallel Phenotype/Genotype Testing PSGT Report – Examples of Discordance17

18. Parallel Phenotype/Genotype Testing PSGT Report – Examples of Discordance18

19. Pol and Integrase Resistance Testing

20. GenoSure PRIme
Monogram’s proprietary Pol assay allows sponsors and investigators to cost-effectively genotype PR/RT and Integrase in a single assay
Please contact Monogram for technical details on this assay

21. Integrase Resistance Testing
Monogram offers both genotyping and phenotyping to assist Integrase resistance in the clinical trial setting

22. HIV Tropism Testing

23. virus entry inhibitors added
The Trofile Assay A.
HIV env expression vector containing patient’s PCR-amplified HIV env fragments
CD4 +
CXCR4 + D. B.
HIV genomic
luciferase vector +
Transfection
Infection E.
virus entry inhibitors added C.
CD4 +
CCR5 + F.

24. Trofile Report
Trofile identifies the tropism of an individual patient's HIV strain – R5, X4, or a combination of these known as dual/mixed (D/M)
Highly accurate, reproducible results using the complete HIV envelope (gp160)
Clinically validated for selecting patients for CCR5 antagonist therapy

25. Trofile DNA
Test Description: Cell-associated DNA isolated from PBMCs is incorporated into Monogram’s proprietary testing vector and run in a single-cycle recombinant assay to determine tropism.
Product Specifications: Analytically validated to identify tropism in cell associated DNA.
Sample Input: Whole blood; PBMC

26. Neutralizing Antibodies/Vaccine Development

27. Monogram Neutralization Assay
gag
pol P R A+  U3
 env
luciferase
U87:
CD4/CCR5/CXCR4
Neutralization Assay:
Incubation with
antibodies or
patient plasma Y
env
luc
a/b
c/d
gp gp41
(patient env gene)
HEK-293
Vectors:
Clone population of viral
env present in plasma,
cell culture, etc.
Drug resistant PR/RT can be used for testing plasmas with RT inhibitors

28. Neutralization Assay Controls
Three control viruses tested with every plasma:
NL43 – neutralization sensitive lab strain
JRCSF – well known primary isolate
aMLV – specificity control:
A test IC50 must >3X IC50 with aMLV to be called positive (green color)
HIV+ Plasma Control:
Tested with every virus in every run
One control for every 7 sera tested
N16, Z23 - Broad, low titered neutralizing capability
Purpose of controls:
Allow Monogram to assess run performance
Allow PI to compare data across time

29. Antibody Neutralization Assay Inhibition Curve
JRCSF
aMLV
Negative curve -
No inhibition of
aMLV by 2G12
Positive curve -
IC50 = 0.56ug/ml
aMLV = specificity control
Anti-HIV antibodies will not inhibit aMLV

30. Assay Formats Multiple formats: HITS 4 dilutions
For screening large numbers of sera
Report % inhibition at each dilution
CURVES
10 dilution curves with IC50, 80, 90, 95
Cutoff for positive is >3X IC50 with aMLV
CURVES PLUS
10 dilution curve and IC50, etc. plus % inhibition at each dilution

31. Other Virology Testing and Services

32. Novel Drug Characterization
Monogram can leverage its virus library and database to construct panels for drug characterization

33. Oncology Testing Services: VeraTag technology and HERmark assay

34. What is a “VeraTag” (VeraTag Reporter)?
How VeraTag Works
What is a “VeraTag” (VeraTag Reporter)?
Three components
Recognition element (most often antibodies)
Cleavable tether (thio-ether or disulfide bond)
Fluoroscein reporters (variable mass-charge)
antigen
antibody
tether
fluoroscein

35. HERmark Assay

36. HERmark Assay Workflow

37. HERmark Report
Classification of HERmark positive, negative, or equivocal
Lab Developed Test
Validated to CLIA standards
Performed in a central, CAP-certified clinical reference laboratory
All cases reviewed by Board Certified Pathologists

38. Additional VeraTag Assays

39. For questions about Monogram technology or assay ordering:
Andrew Gale
Director, Pharma Collaborations