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Integration of Imaged cIEF and MS with a Novel Microfluidic System for Real-Time Analysis of Biologics
Erik Gentalen, Chief Technology Officer
WCBP, January 30, 2019
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TRANSFORMATIVE SOLUTIONS for accelerating biotherapeutics
Comprehensive Biologic Product Quality Analysis
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Comprehensive Biologic Product Quality Analysis
100x Higher Throughput
100x More Cost Efficient
◆16 team members; Newark, CA
◆$9.5M Series A, Oct 2018
$15M+ total financing and grants
◆7 Early Access Pharma Customers
Bayer, Merck, Genentech, Celgene, MilliporeSigma
◆Core Technology Patent Issued
◆Commercial Launch mid-2020
Launch Demo Lab & Tech Evaluation, mid-2019
Success Innovating at the Intersection of CE, Microfluidics, MS
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Lou Dietz, M.S.Engineering/Systems Development
Khushroo Gandhi, Ph.D. MEMS Fabrication, surface chemistry
Anne Kopf-Sill, Ph.D. Microfluidics/Product Development
Bill Hyun, Ph.D. Analytical instrument Development
Lena Wu, Ph.D.CEO & President, Co-founder
Erik GentalenCTO, Founder & InventorPatent Agent
Wally Ausserer, Ph.DVP, Corp Dev & Marketing
Ian Walton, M.E.VP, Product Development
Don Arnold, Ph.D.Microfluidics, MS Interface Eksigent
Technical advisors with success in product development and commercialization
12M+ Financing
$2.7M Grant Funding
Growth in biologics has created a crisis in
product quality characterization
◆ Explosive growth in $200B
Biologics market
◆ Emerging biosimilars market to
reach $30B in 2020
◆ Bioproduction is complex and
requires constant testing
◆ Compressed timelines under FDA
Breakthrough Therapy
Designation
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Confidential
Blaze Critical at Every Stage of $7B Biologics Production Market
Bio
Blaze System: Instrument, Software, Microchip, Reagents
• Blaze Instrument
– Bench-top
– Stand-alone (QC mode) or direct interface to MS (Analytical mode)
– Interfaces with multiple MS brands/
models
– 96-well plate with autosampler;
unattended operation
– 280 nm absorbance; whole column
detection
– Software/Analysis
¡ Automated pI calibration; Peak
correlation with mass spectra; Data export
• Charge Heterogeneity Assay Kit, 100
samples
– 1 Microchip per kit
– Reagents for 100 samples
– pI standards
Blaze Instrument
Microchip/cartridge & reagents
Integrated peak software analysis
Blaze System
Customer’sMass Spec
R&D GMP
Cell BankingDownstreamProcess Dev
Formulation
Mftg
CommercialManufact(10+ yrs)
• BioPharma Development • CMOs• Commercial Manufacturing• Biosimilars
Upstream Process Dev &
Characteriz
Cell Line Dev
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Rapid, HT Product quality characterization
Collect fractions
Scale up
LC Electrospray
Purify protein
Molecular mass identifies peak
• What is this
other peak? • How has the
protein been
modified?
Method Development
Tris
NaCl
pHAcetate
MES
Detection by cIEF
CharacterizationBy MS
Blaze seamlessly
integratesto the customer’s MS
Current workflow
Low capacity and months per analysis
Separate by IEX Chromatography
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Intabio’s Blaze Streamlines and Accelerates Product Quality Analysis
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Blaze
Proprietary Microfluidic Technology Integrates Key Analytical Functions
15-20 min analysis time per sample
• cIEF separation• Real-time, UV absorbance imaging
• MS sample prep • Peak mobilization
Mass spec + + + +
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Sample Introduction
Mass Spec
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Electrolyte
• Sample delivery by electrospray to MS
Separation and Imaging column, 50 mm length
Electrospray Tip
Separation Channel
ChannelTerminus
Microfluidic Chip
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Core Technology Patent Issued Feb 2019
CONFIDENTIAL
Blaze Peak Resolution is Comparable to Commercial cIEF Systems
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• pI Standards imaged in real-time during 10 minute separation on Blaze microchip
• Marker concentration, 12.5 mg/mL
• Separation of pI marker standards ladder is linear
pI 3.38
pI 4.05pI 7.00
pI 8.40
pI 9.99
pI 10.17
Blaze System Imaged cIEF Analysis of pI Markers
Blaze System NIST mAb Profile Meets Industry Standards
2 Lys
1 Lys
(Main) 0 Lys
Acidic
1.5% Pharmalyte 3 to 10, 1.5% Pharmalyte 8 to 10.5, 20 mM Arginine and 250 µg/mL NISTmAb in DI H2O
(Main) 0 Lys
1 Lys
2 Lys
Acidic
ProteinSimple Technical Seminar, CE Pharm 2016
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Blaze System Imaged cIEF Analysis
Chip 2811
Chip 2808
Chip 2805
pI 7.0pI 10.0
1 Lys
Main (0 Lys)
Acidic
2 Lys
NIST mAb Profile is Reproducible Across Multiple Chips
1.5% Pharmalyte 5 to 8, 1.5% Pharmalyte 8 to 10.5, 20 mM Arginine and 250 µg/mL NISTmAb in DI H2O
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Charge Heterogeneity Analysis of Model mAb
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Blaze System Imaged cIEF Analysis
• Separation comparable to conventional cIEF systems
• Assay optimized for mass spec compatible reagents, e.g. formamide replaces Urea
Trastuzumab
Trastuzumab by imaged capillary cIEFDai et al Anal Chem 2018
BMS Analysis
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Novel Demonstration of High Resolution cIEF Peak MobilizationReal-Time Imaging of Trastuzumab During Focusing and Mobilization
At 2.5 min, initial phase of focusing At 6 min, focusing completed
At 8.5min, mobilization of peaks toward the ESI tip is in progress
At 9.5 min, trastuzumab peaks are about to enter the ESI tip. Note the nearly baseline
resolution achieved between the two main peaks for trastuzumab
Trastuzumab
pI 7.00
pI 9.99
Trastuzumab
pI 7.00
pI 9.99
Trastuzumab
pI 7.00
Blaze Microchip Electrospray Sensitivity Enables Detection of NIST-mAb Isoforms
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• NIST mAb infusion, 500 mg/ml
• Stable Taylor cone and MS ion chromatogram
• Demonstration of nanoflow (~100nL/min) for electrospray ionization
• Bruker Compact QTOF
ESI Tip
Chip MS
Top view of microchip electrospray delivery into an adjacent MS 16/18 Glycoforms Detected
Mass Spectra
Ion Chromatogram
Deconvoluted Data
Intabio Demo Lab and Technology Evaluation Service
• Early Access Program: demo lab and sample analysis service
o Visit for hands-on Blaze demo and MS integration (multiple MS systems)
o Send samples to Intabio for analysis
• Blaze 1.0 System commercial launch mid-2020
o Smaller footprint, internal autosampler, 21 CFR part11 ready software, consumable chip kit.
• Sign-up and additional details coming soon www.Intabio.com
• Based in Newark, California
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Blaze Early Access System
THANK YOUwww.Intabio.com
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Comprehensive Biologic Product Quality Analysis