1. “For over 24 years, we’ve worked hand in hand with industry,
academic, and government researchers to help solve difficult
assay development and protein quantification problems.”
Stephen Parent, PhD
Director of Business Development
sparent@pblbio.com
(732) 777-9123 x117
2. PBL Assay Science History
• Founded in 1990 by Dr. Sidney Pestka
• 35 employees based in Piscataway, NJ
• World’s largest producer of interferons and related
cytokine products for the research market
– ELISA and multiplex assay kits
– Recombinant proteins
– Antibodies
– Assay services
– Custom services
3. High Quality Assay Solutions Interrogating
Interferon and Cytokine Biology
4. Assay Solutions Interrogating IFN and Cytokine Biology
• Off the Shelf Assays
– More than 70 Interferon ELISAs, antibodies and cell-
based assays
• Assay Services Offerings
– Sample Testing and Screening
– More than 40 Singulex Erenna biomarker assays
– Assay Design and Development
– Custom Services
– We provide ~ 70 different services
5. Assay Services Capabilities
Cell-Based Assay Development
Antiviral Assays
Cytokine/Chemokine Bioactivity Assays
Immunoassays
Development
Single Analyte & Multiplex ELISAs
Ultrasensitive Cytokine Detection Assays
using Singulex Erenna® technology
More than 40 Assays Available
Human and Mouse
Reagent Development & Characterization
Assay Optimization
Technology Transfer
Consulting in Assay Development and
Therapeutics Development
6. IL-17
Ultrasensitive Cytokine Detection Service
using Singulex Erenna® Immunoassay System
Modified IA process
using magnetic beads
Single Molecule
Detection (SMD)
technology
50- 1000 fold higher sensitivity over existing IA
products
More than 40 Assays Available
7. IL-17
Singulex Single Molecule Counting (SMC)
Erenna®
4.
5.
► Elution buffer
dissociates the
complex (analyte +
detection antibody
in simple buffer)
► Transfer to 384-
well plate
► Draw eluate into the
instrument for reading
► Fluorescently labeled
antibodies introduced
to laser confocal
microscope digital
counter
► Molecules are
individually detected
and digitized
Elution Molecule counting
Proprietary Singulex technology
Sample collection
2.
3.
► Transfer samples
to 96-well plate
► Incubate with
capture magnetic
antibody-coated
beads
► Wash to remove
unbound
antibodies
► Incubate with dye-
labeled detection
antibodies
1.
► Low abundance of
certain biomarkers in
complex biological
samples
Antibody incubation Washing & fluorescent labeling
Standard immunoassay protocol
More than 40 Assays Available
9. Examples of Available Cell-based IFN Assays
Cytopathic Effect (CPE) Inhibition Assay
Neutralization of 1 U of IFN in
cytopathic effect (CPE) inhibition assay
A549 human lung cancer cell line with
encephalomyocarditis virus (EMCV)
Cell-based Neutralizing Antibody Assay
10. Examples of Cell Proliferation & Anti-Proliferation Assays
OVCAR-3 human adenocarcinoma line TF-1 human erythroleukemia line
Proliferation quantified using soluble tetrazolium reagent
11. Immunoassay Products
• 10 Human Interferon ELISAs
• 3 Mouse Interferon ELISAs
• 1 Cynomolgus/Rhesus Interferon ELISA
• 1 Human soluble IFNAR2 ELISA
• 2 Multiplex ELISAs quantifying Type I, II and III IFNs
and other cytokines
• Custom Multiplex ELISAs are also possible
12. You can have confidence in PBL ELISAs!
• Human IFN alpha ELISA study (2008)
• Split samples run at a CRO and PBL
• Slope = 0.991
• Correlation coefficient = 0.9931
• Human IFN beta ELISA study (2009)
• Split samples run at a CRO and PBL
• Slope = 1.036
• Correlation coefficient = 0.9966
• Human IFN beta ELISA study (2014)
• P. Conliffee et al. AAPS Biotechnology Conference
• Validated analytical range 2.34 - 150 pg/ml
• Dilutional linearity up to 1 : 2187
• Interassay precision (CV) ≤13.1%
• LLOQ 2.34 pg/ml
• No IFN-b detected in normal sera.
• Median quantifiable IFN-b was 17.3 pg/ml in MS sera (n=47), with 26
quantifiable & 21 below LLOQ.
• MS samples across different sites showed excellent correlation (r2=0.9975)
13. VeriKine Human Interferon ELISA Kits
• Human IFNa Multisubtype Serum Kit
• Validated at numerous sites
• Kit standard calibrated to NIH human IFN alpha
standard
• Dual detection range
– 12.5 – 1000 pg/ml
• LLOD: 1.6 pg/ml; LLOQ: 5.7 pg/ml
• Intra-assay, Inter-assay, and Inter-lot CV < 8%
• Compatible with 50% normal human serum, 50%
plasma, TCM and nasal lavage
• Unique broad subtype cross reactivity
• Detects Intron-A®, Roferon®, pegylated interferon alphas
• Human Interferon Beta Serum Kit
• Developed to meet ICH guidelines on validation of
analytical procedures
• Validated at 3 independent labs to date
• Kit standard calibrated to NIH human IFN beta
standard (correlates with NIBSC standard)
• The most sensitive human IFN beta ELISA in the
market
• Detection range: 1.2 - 150 pg/ml
• LLOD: 0.3 pg/ml; LLOQ: ~2 pg/ml
• Intra-assay, Inter-assay, and Inter-lot CV < 8%
• Compatible with normal human serum, plasma,
tissue culture media
• Characterized on MS patient sera
• Detects Betaseron®, Avonex®, Cynomolgus,
Rhesus IFN beta
• Detects IFN beta bound to soluble IFNAR2
receptors
14. VeriKine-HS High Sensitivity
Human Interferon Beta Serum ELISA Kit
90% of Patients on IFN
Beta therapy have
quantifiable IFN Beta
2% of Healthy Donors have
1-2 pg/ml IFN-Beta
No elevation of IFN Beta
on other therapies (p=0.4)
IFN Beta in Normal and MS Patient Sera
Detected by ELISA
IFN
Beta
Treated
n=29
N
o
IFN
Treatm
entn-25
N
H
S
n=193
0
10
20
30
40
50
60
70
80
90
100
110
120
IFN-Beta(pg/ml)
16. PBL’s New Human IFNa All Subtype ELISA (Cat. 41115)
is the most sensitive assay kit on the market
Mean LLOD &
LLOQ values on
ALL 12 human
IFN-α subtypes
LLOD & LLOQ values
from IFN-α2 alone;
subtype quantitation
information available
LLOD & LLOQ values from IFN-α2 alone.
No subtype quantitation information available.
*Competitor data obtained from public sources,
provided for comparison only.
0
2
4
6
8
10
12
PBL 41115 PBL 41110 Competitor A Competitor B Competitor C Competitor D(non-
ELISA technology)
LLOD (pg/ml) LLOQ (pg/ml)
18. ELISA assays for soluble Type I and Type II IFN receptors
Soluble Type I and Type II Receptors are elevated in Males
19. sIFNAR2 ELISA Assay
sIFNAR2 Levels in Autoimmunity
Normal n=59
SLE n=111
RA n=16
Sjogren's n=11
Scleroderma n=10
0
2500
5000
7500
10000
10000
35000
p=0.008
sIFNAR2pg/ml
20. Reagent Products
• Proteins
– Human Interferons
– Mouse Interferons
– Rat Interferons
– Other Interferons
– Human Cytokines & Growth
Factors expressed in Human Cells
• Polyclonal Antibodies
– PAbs to Human Interferons
– PAbs to Mouse Interferons
– PAbs to Rat Interferons
– Labeled PAbs
• Monoclonal Antibodies
– MAbs to Human Interferons
– MAbs to Mouse Interferons
– MAbs to Rat Interferons
– MAbs to Porcine Interferons
– Labeled MAbs
21. PBL Assay Science Team
PRaDAS Quality Control
Tom Lavoie Sehrish Ajmal*
Taher Fatakdawala* Karen Zipf*
Robert Niemczyk* Production Team
Xiao-Hong Lin* Daniela Rotaru
Tara Stauffer* Karen Murphy
Michael Skawinski* Rocio Acosta
Lara Izotova* Ksenia Makarenko
Eric Zhu* Natasha Kugakolova
Barbara Schwartz
Nirupama Putcha Collaborators
Sanjna Sanghvi Ray Donnelly (FDA/NIH)
Katherine Carrino-Bednarski Phyllis Conliffe (Smithers Avanza)
Jonathan Ferreira Ira DuBey (Smithers Avanza)
