MyCartis technology intro

From an idea to the Evalution™ product

EPFL – SV (for students of class on Reg. medicine/Microfluidic based Dx)

20. Oct. 2014

Didier Falconnet, dfalconnet@mycartis.net, www.mycartis.net (/blog)

Context

• Rudi Pauwels, PhD in pharmacology and life-sciences star-entrepreneur on sabbatical in the lab of Philippe Renaud (EPFL)

• Can we leverage microtechnologies to develop better diagnostic tools?

• 2007 Rudi Pauwels, Philippe Renaud and Nader Donzel found Biocartis S.A. (EPFL, PSE)

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The ambition: Making healthcare personalized

• Diagnosing patients at the molecular level (beyond current test method)

• Diagnosis of complex diseases and response to treatments are often associated with multiple biomarkers with time-dependent signatures

• Multiplex (multiple biomarkers simultaneously, high information density)

• Rapid and cost effective (time to results, cost per data point)

• Easy to use technology yet flexible (limited skills required)

• Quantitative (measuring the response of a treatment for fine tuning the

dose – one example of personalized medicine)

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*DeciBio market report Feb. 2012

Strong development of Dx technologies

• IVD spent currently represents <1% of total WW healthcare spent (~$5.5T+), although results from these tests account for 60-70% of all treatment decisions*.

• The “one fits all” drug and dose is over

• Today, strong focus on diagnostics as it is a corner stone of personalized medicine

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From start-up to SME in just a few years

• 2007 Biocartis is founded on seed money (R. Pauwels) and develops initial concepts and demonstrations

• 2010 Biocartis (< 10 employees) acquires the “Apollo” program from Philips with a new round of investments

• 2014 Biocartis (~ 160 employees in CH and B), launch of the Idylla™ platform (PCR based, sample-in data-out)

Mycartis spins-out (~ 50 employees in CH and B, integration of Pronota NV) and launch Evalution™ platform (Multiplex biomarker analysis, RUO)

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Historically 2 platforms in the same company

• Idylla™ − Diagnostic samples in/result

out platform based on real time PCR

• Evalution™ − Multiplexed biomarker analysis

platform for research (and diagnostics)

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Requirements

• Multiplex

• Rapid

• Easy to use technology yet flexible

• Quantitative

+

• Sensitive and reproducible/robust

• Versatile

• Facilitating development of multiplexed assays

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Multiplexing for “rich data”

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• Traditional technologies measure one molecule per reaction

• Multiplexing enables measuring multiple molecules simultaneously

• Increased information density per sample volume

• A variety of multiplexing technologies have emerged in the past decade, each with its encoding strategy and limitations

Innovative solution for multiplexing

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Functionalize with capture molecules

Aliquots to create master mix

Add master mix to microfluidic cartridge

• Our starting point was IP from memobead

• Barcoded fluorescent beads for multiplexed analysis

• Concept demonstrated but impractical for industrialization

• Development of a novel digitally encoded particles (foundation of MyCartis technology)

Evalution™ animation

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Digitally encoded microparticles

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Production process from state-of-the-art semi-conductor industry

Digital encoding yields thousands of codes

Compatible with well-established surface chemistries

Activated of bio-functionalized microparticles

High reproducibility from monocristalline silicon

Ideal flat surface for 2D imaging

Aggregation of 1800+ pixels per single microparticle

Microparticle for high data quality

Fluorescence detection zone • >1800 pixels of data per microparticle • Uniform signal (2D surface) • No cross-talk between particles • Smart fluorescence data aggregation

Robust digital coding • Unambiguous • Stable • Quality control integrated • Simple to produce • On-the-fly decoding

Clean and well characterized materials • Crystalline silicon matrix • Well established and robust chemistry • High quality production processes

Microtechnology enabled surface features • Optical signal enhancement layers • Flow enhancing structures

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Evalution™ core components

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Encoded microparticles

Microfluidic assay plate

Microfluidic assay plate When high technology meets simplicity

Channel cross-section 400μm x 17μm

High-fidelity replication for planar arrangement of

microparticles ̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶ μm tolerance ̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶

Based on titerplate standard for full compatibility with

lab automation

Run up to 16 assays in batch or on different dates

Capacity of 150-plex per channel

Optimized environment for fast, reproducible, and

specific bindings

Getting more from less in miniaturized environment ̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶ μL range ̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶̶

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Simple and robust cartridge, no moving parts (i.e. no valves)

Enables mass production by plastic injection

Particles are retained and allow repeated imaging of the reaction chamber

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Automated data sampling

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Visual aspect and software

Data are displayed real-time upon imaging

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Instrumentation

Bench top instrument with automated controls, separate microparticle loading station

Green laser optics for ultra sensitive and low noise fluorescence imaging

High pressure operation for robust fluidics

Three independent temperature zones from 25 to 95 °C for assay flexibility

Independent operation of the 16 reaction chambers

Real-time or end-point imaging

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Evalution is composed of 4 subsystems

• Optical: x-y-z scanning objective, 532 nm laser, CMOS cam. Bright-field and Fluorescence mode.

• Thermal: 3 independent temperature zones (25-95°C)

• Fluidic: common mode pressure with up to 2000 mbar differential pressure

• Software: real-time data analysis and dynamic control over assay environment

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Microfluidics for short assay times and quantitative results

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• Reaction limited regime (biophysical ideal conditions)

• Not governed by diffusion

• Fluorescence dependent of concentration (in RLR)

• Short assay times minimized non-specific binding

Reaction limited regime

Mass transfer limited regime

Squires, Messinger and Manalis. Nat. Bio. 2008

Flow rate function of tot. #particles and DP

• Data for 500mbars and 25°C in PBST

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1 mm loaded : 250 particles

• And many more features but outside scope of this presentation

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Summary on technology intro

• Novel solution for biomarker analysis

− Multiplex robust encoding

− Flexible comp. with different workflows, dynamic control over assay env.

− Versatile different sample and analyte types

− Adaptable no batch required, short time to result

− Easy to use visual, real-time, no plate transfer, only 1 instrument

− Fast reaction limited regime in microfluidic channels

− Quantitative sensitive fluorescent readout

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Looking back to the start…

• A (good) idea that solves a question, a problem or opens possibilities not though of before

• Be open and curious across disciplines, network

• Change is good !

• Be prepared for ups and downs, be flexible

• Team-up with the “right” people, identify trends

• We are now commercially launching our product with a CEO knowing the market very well.

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Outlook

• Launch on Research market (now)

• IVD

• POC

• Matching sensitivities of digital systems

• Further simplifying workflow for super and hyperplexing

• Menu expansion for different disease areas (with partners)

• www.mycartis.net (/blog)

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Thank you

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Flexible workflow and readout mode

• Traditional multiplex sandwich −endpoint (min. 5 pip. steps)

• Simple and fast coflow sample and det. Ab −kinetic or endpoint (1 pip. step)

• Post-PCR detection −kinetic or endpoint (1-3 pip. steps)

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Benefits of Evalution™ (and coflow)

• Labour − You save multiple pipetting steps

− No need to transfer plate between instruments

• Time − Great reduction of assay time

− Great reduction of hands on time

• Workspace − 1 instrument versus 3 instruments

• Reagents − Evalution uses differential air pressure for media transport

− No drive or sheath fluid needed

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Immunoassay workflow drastically accelerated

− Simple workflow with minimal hands-on-time

− Real-time or end-point readout

− Identical workflow with increased plex-size

TAT: < 30 min

HOT: <10 min

5 pipet steps

Evalution

Seq Flow

TAT: 10 min

HOT: <5 min

2 pipet steps

TAT: > 2.5 h HOT: > 1h >15 pipet steps

ELISA platform

Incubate Read Preparation Incubate Wash Wash Incubate

0 60 120 180 240 min

TAT Turn Around Time HOT Hands-On Time

*Preparation for ELISA

includes performing multiple dilutions to enter dynamic range

*

Evalution

Co Flow

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Thermal subsystem:

• 3 different and independent temperature zones

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Inlet zone

Denaturation zone

Detection zone

Prepare the reagents

• Couple capture molecules • Create a mix

1

Prepare the assay plate

• Loading of pre-mixed encoded microparticles

• Multiple channels at a time

2

Insert the plate

• Select assay parameters

• Edit/load protocol 3

Run the assay

• Load the sample • Monitor the reaction and report results

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The workflow, 2 protocols

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Full flexibility for user-defined assay

Fast workflow with pre-filled assay plate