Monogram Biosciences
Pharma Collaborations
-A LabCorp Center of Excellence –
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
Monogram Biosciences
A LabCorp Center of Excellence
LabCorp’s Platform
220,000 customers 31,000 employees
1,500 patient centers 440,000 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.
Monogram’s Technologies are Offered Directly at
Monogram and through Esoterix Clinical Trials
Services, a Division of LabCorp
Central Labs
Dedicated labs for
clinical trials
Cranford, NJ
Mechelen, Belgium
Beijing, China
Singapore
Broad, validated
test menu
Bioanalytical
Labs
Biomarker
Discovery
Companion
Diagnostics
8 Centers of Excellence
Small and large molecule
Discovery and development
Discovery, development,
commercialization
Coagulation
Endocrinology
Oncology
Infectious disease
Anatomic pathology
Molecular pathology
Flow cytometry
Method development
and validation
GLP biomarker services
Esoteric Testing
FDA experience
MS-based assays
IVD Trial Participation
LC/MS/MS
Non-GLP discovery
services
Cell-based assays
Global expertise:
Cranford, NJ
Mechelen, Belgium
San Diego, CA
West Trenton, NJ
Successful PMA submission
40,000 ft2 biorepository
Test method
harmonization
Commitment
to Quality
Integrated Data
Management
Dedicated
Project
Management
Testing Services Across all Phases
of Development
Discovery
• Screening drug candidates
• Characterizing resistance
Preclinical
Clinical
• Screening patients for enrollment
• Optimizing background therapy
• Characterizing treatment failure
Post-Marketing
• Defining clinical cut-offs
• Enhancing market positioning
HCV Testing Services
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
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
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)
HIV PR/RT Resistance Testing
PhenoSense HIV Cell Assay
Resistance Test Vector
DNA
A-MLV env
DNA
+
Transfection
Infection
293-A
Pseudovirions
PR inhibitors added
RT inhibitors added
PhenoSense HIV Data
Patient virus exhibits reduced susceptibility to the drug
Patient:
Control:
GenoSure MG Report
Monogram’s GenoSure MG
report combines LabCorp’s
genotyping capabilities (for
faster turnaround), and
Monogram’s proprietary
algorithm…
Parallel Phenotype/Genotype Testing
PhenoSense GT Assay (PSGT)
Phenotype
Patient virus
RT-PCR
Genotype
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
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
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
Phenotyping
• Direct measure of susceptibility
• Ease of interpretation
• Demonstrates effect of complex
combinations of mutations
• Resistance and Cross-Resistance
• Re-sensitization
• Partial Activity
• Replication Capacity
• Immature algorithms for new
agents
Parallel Phenotype/Genotype Testing
PSGT Report – Examples of Discordance
Parallel Phenotype/Genotype
Testing
PSGT Report – Examples of Discordance
Pol and Integrase Resistance Testing
GenoSure PRIme
Monogram’s proprietary Pol assay allows sponsors and investigators to costeffectively genotype PR/RT and Integrase in a single assay
Please contact Monogram for technical details on this assay
Integrase Resistance Testing
Monogram offers both genotyping and
phenotyping to assist Integrase
resistance in the clinical trial setting
HIV Tropism Testing
The Trofile Assay
A.
HIV env expression vector
containing patient’s PCRamplified HIV env fragments
B.
HIV genomic
luciferase vector
CD4 +
CXCR4 +
D.
+
Transfection
Infection
E.
C.
virus entry
inhibitors
added
F.
CD4 +
CCR5 +
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
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
Neutralizing Antibodies/Vaccine
Development
Monogram Neutralization Assay
Vectors:
Drug resistant PR/RT can be used for testing plasmas with RT inhibitors
gag
P R U5
P
Clone population of viral
env present in plasma,
cell culture, etc.
P
pol

U3
luciferase
gp120
a/b
Neutralization Assay:
gag
R A+
 env
pol
gp41 A+
c/d
(patient env gene)
luc
env
Y
HEK-293
Y
Incubation with
antibodies or
patient plasma
U87:
CD4/CCR5/CXCR4
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
Antibody Neutralization Assay
Inhibition Curve
JRCSF
Positive curve IC50 = 0.56ug/ml
aMLV
Negative curve No inhibition of
aMLV by 2G12
aMLV = specificity control
Anti-HIV antibodies will not inhibit aMLV
Assay Formats
Multiple formats:
1. HITS
– 4 dilutions
– For screening large numbers of sera
– Report % inhibition at each dilution
–
2. CURVES
– 10 dilution curves with IC50, 80, 90, 95
– Cutoff for positive is >3X IC50 with aMLV
3. CURVES PLUS
– 10 dilution curve and IC50, etc. plus % inhibition at each
dilution
Other Virology Testing and Services
Novel Drug Characterization
Panel #
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
Monogram can
leverage its virus
library and database
to construct panels
for drug
characterization
AMP
14.6
10.6
13.2
5.1
6.3
13.1
8.9
2.1
12.2
5.5
28.8
4.2
17.7
10.4
31.6
16.7
48.8
20.5
2.8
29.6
7.7
42.1
4.9
9.0
3.3
14.6
7.0
130.0
13.8
7.0
10.1
22.2
14.3
40.9
11.2
12.5
4.9
42.5
4.9
15.0
14.1
28.2
4.2
15.3
71.8
IDV
250.0
46.9
45.0
48.1
31.1
11.9
10.6
23.6
39.5
151.2
250.0
82.6
35.8
40.6
58.6
11.2
14.5
20.3
169.2
250.0
27.7
16.9
23.2
15.0
10.2
38.3
35.9
22.5
40.8
12.7
20.2
55.1
25.5
28.7
250.0
6.7
12.3
64.0
10.6
11.4
21.5
20.5
18.1
20.5
21.3
NFV
148.7
109.8
50.1
51.9
21.0
21.5
35.1
23.1
55.8
91.0
74.2
98.1
93.8
97.7
85.0
53.5
32.5
30.1
167.2
297.4
20.0
50.1
41.5
30.6
39.4
55.9
64.7
29.4
58.2
30.5
40.1
185.9
54.5
100.7
155.0
18.5
13.1
215.6
24.4
52.0
74.2
69.3
34.1
18.4
104.3
RTV
275.0
275.0
28.5
123.7
79.2
141.9
191.5
42.2
29.0
79.9
275.0
116.5
275.0
78.1
275.0
275.0
275.0
275.0
89.2
275.0
107.6
275.0
59.5
169.2
40.2
22.8
44.4
275.0
132.9
87.3
80.0
275.0
217.4
63.5
275.0
60.1
81.7
275.0
54.2
42.4
66.1
149.2
275.0
85.7
61.7
SQV
96.5
58.4
61.3
39.7
20.0
15.2
83.2
143.1
52.9
561.7
550.0
596.6
33.8
113.7
550.0
69.0
36.3
27.4
550.0
550.0
4.9
127.6
61.3
53.6
1.0
24.1
96.7
550.0
94.4
46.6
74.5
226.2
68.6
96.1
550.0
9.5
25.4
348.2
68.2
126.7
170.2
101.6
41.2
22.8
77.3
LPV
201.2
58.7
11.6
nt
nt
49.6
nt
nt
nt
60.8
148.8
nt
68.4
68.9
nt
nt
nt
97.1
72.1
156.0
22.3
93.0
nt
nt
6.1
nt
13.8
273.1
67.3
38.2
nt
nt
nt
66.5
nt
51.5
nt
149.6
nt
nt
15.6
65.2
54.2
45.2
43.9
L10 K20 L24 D30 V32 L33 M36 M46 I47 G48 I50 F53 I54 A71 G73 V77 V82 I84 N88 L90
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Oncology Testing Services:
VeraTag technology and
HERmark assay
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)
O
antigen
HOOC
S
O
tether
OH
fluoroscein
O
O
H
N
antibody
O
N
H
NH
HERmark Assay
HERmark Assay Workflow
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
Additional VeraTag Assays
For questions about Monogram technology or assay ordering:
Andrew Gale
Director, Pharma Collaborations
agale@monogrambio.com