Cedex Analyzers for Process Control in Bio-Processing · Common applications of monoclonal mouse...

Cedex Analyzers for Process Controlin Bio-ProcessingJune 28st 2018

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Webinar agenda

1. Cedex bioprocess control solutions Roche CustomBiotech

2. Application of new Mouse IgG test Roche Diagnostics

3. Application of new Osmolality estimation Roche Pharma

4. Improved solutions for analytics workflow control AGU Industrial ITand data management

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Webinar agenda





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Roche CustomBiotechKnow-how and products to power your ideas

Customers worldwide in the pharmaceutical and diagnostics industries

Roche Pharma Roche Diagnostics

RochePharma Genentech Chugai

Centralized and PoC Solutions

Molecular Solutions

DiabetesCare

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Roche CustomBiotech in Penzberg, GermanyAdvantage of Pharma and Diagnostics on one campus

Germany

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Roche CustomBiotech in Penzberg, GermanyAdvantage of Pharma and Diagnostics on one campus

Shared research facilities, technologies and discoveries

RochePharma

Process development,bio-manufacturing

RocheDiagnosticsBroad expertise

in analytics

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Roche CustomBiotechSolutions for all steps in bio-processing

Development

Cell isolation Cell yield &viability testing

Cell expansion, media optimization

Passaging & harvesting

LiberasePortfolio

Rec Trypsin, GMP

Cedex Bio Analyzers

Cedex HiResAnalyzer

Manufacturing

Scale-up

Liberase blends, GMP

Recombinant Trypsin, GMP

CedexAnalyzers

MycoTOOL kit

QC testing

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Roche CustomBiotechInstrumentation for cell culture process control

Substrate, metabolite & product monitoring Cell density & viability

Cedex Bio Analyzer

Cedex HiRes Analyzer

Cedex Bio HT Analyzer

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Cedex HiRes AnalyzerImage-based cell counter

Cell density

Cell viability

Cell morphology

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Cedex HiRes AnalyzerHigh-resolution image capturing

Image recognitionResolution

live cell particledead cell aggregate

Algorithms for differentiation of

• viable cells dead cells

• cell aggregates other particles

• 0.8 μm/pixel• Suited for cell diameters

of 2 – 40 μm

17 pixels

14 μm cell diameter

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Cedex Bio / HT AnalyzerSame tests for different sample load

Cedex Bio Analyzer Cedex Bio HT Analyzer

Throughput ≈ 75 tests/h 250 – 400 tests/h

Sample capacity 8 samples at once, 90 samples at once,

Parallelization

Auto-dilution selectable free programmable

- Continuous loading during operation -

- up to 60 tests -

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Cedex Bio AnalyzerRobustness based on long-established Roche technology

Track record of a Cedex Bio Analyzer in a large-scale fermentation lab of Roche Pharma:

in operation for 8 years now

> 125 000 tests performed

only 8 cases of instrument outage

5 of 8 failures could be fixed at once, total down time <10 days

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Ala-Gln (GlutaMAX)

Cholesterol Calcium Galactose Glucose

Glutamate Glutamine Glycerol Iron Magnesium

Acetate Ammonia Ethanol

IgG (human) Mouse IgG

Substrates

Metabolites

Products

Formate Lactate LDH

Optical Density Total Protein

Phosphate Potassium Pyruvate Sodium Sucrose

Osmolality, calc.

Cedex Bio test menuFor process control in cell culture and microbial fermentation

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20152013 20142011 / 2012

Iron

Phosphate

Magnesium

Calcium

Acetate

Glycerol

Total Protein

Improved Multi-Controls

Cholesterol

Pyruvate

Galactose

Sucrose

Glutamate V2

Glutamine V2

Acetate V2

System launchesCedex Bio / Cedex Bio HT

2016

Optical Density(OD)

Formate

Ethanol

2017

Ala-Gln(GlutaMAX)

2018 ff

…

…

…

Galactose V2

Mouse IgG

Osmolality calculated

Cedex Bio test menuContinuous menu extension according to the needs in bioprocessing

Glucose Glutamate Glutamine IgG Lactate LDH NH3 Potassium Sodium

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Webinar agenda





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New Mouse IgG testCommon applications of monoclonal mouse IgG antibodies

Research

Immunoassays (e.g. ELISA, Western blots, fluorescence microscopy)

In-vitro diagnosticsImmunoassays (e.g. Clin Chem, flow cytometry, immunohistology)

PharmaDrug discovery phase for pharmaceutical antibodies

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New Mouse IgG testProcess control in mouse IgG production

So far, determination of product titer was done by:

• Manual methodslike manual ELISA, OD-280, Biuret)

• Centralized instrumentationlike HPLC or other types of analyzers

- poor reproducibility- excessive hands-on time

- long time to receive results- dependence on other departments

Now available:

• Cedex test

- automated testing

- high reproducibility

- results after 10 – 15 min

- wide test range

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New Mouse IgG testAssay characteristics

Launched in May 2018

Specificity: Fc part of mouse IgG, subtypes IgG1, IgG2a, IgG2b

Range: 10 – 7,500 mg/L

Generic calibrator and controls provided, option for custom calibration

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New Mouse IgG testAssay verification in Roche Diagnostics manufacturing

Linearity of the assay:

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New Mouse IgG testAssay verification in Roche Diagnostics manufacturing

Accuracy with all IgG subtypes (compared to validated Elecsys test):

IgG1 IgG2a IgG2b

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Webinar agenda





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Osmolality in bio-process control

Purpose

• Quality control of media and supplements, general check for correct water content

• Continuous monitoring of osmotic pressure in cell cultures,to avoid critical dilution or concentration of the medium

Common method

• Determination of freezing point depressionusing a special osmometer instrument

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Cedex OsmolalityCalculation based on relevant cell culture media components

Osmolality

Na+ Cl- > 85%

Glc Gln Lac- 3 – 12%

K+ Ca+ Mg+ NH4+ HCO2- SO42- 3 – 8%

Amino acids Proteins Others < 3%

Cedex profile:

Determination of Na+ + Glc + Gln only

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Cedex OsmolalityCalculation based on relevant cell culture media components

Osmolality

Na+ Cl- > 85%

Glc Gln Lac- 3 – 12%

K+ Ca+ Mg+ NH4+ HCO2- SO42- 3 – 8%

Amino acids Proteins Others < 3%

Cedex profile:

Osmo = (2 * Na+ + 1.3 * Glc + Gln) * 1.003

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Cedex OsmolalityPerformance evaluation

± 10 % range

Comparison “freezing point“ vs. Cedex calculation

• 5,800 cell culture samples, 30 different culture media

• Accuracy: 98 % of results in ± 10 % range

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Cedex OsmolalityPerformance evaluation

Comparison “freezing point“ vs. Cedex calculation

• Examples of two CHO cell cultureswith different feeding principles

• Samples were taken daily over thefermentation period

• Accuracy: < 10 % deviation

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Osmolality Cedex Bio HTMethod evaluation in Pharma Operations

• 250 samples of CHO fermentation runs, 5 different clones, chemically defined media

• Freezing point depression as reference method

• Sodium, glucose and glutamine measured on Cedex Bio HT

• Adjusted empiric formula used

• Comparability and correlation of the two methods are tested

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Osmolality Cedex Bio HTComparability (Clone 1)

Comparability• General equivalence to reference

method (on average 96% recovery)

• Both methods show similar course

• Calculation with Cedex Bio HT lower as reference method

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Comparability• General equivalence to reference



• Calculation with Cedex Bio HT lower as reference method

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Comparability• Very good equivalence to reference



• No offset between methods

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• No offset between methods

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• Both methods show same course

• No offset, almost same results

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Osmolality Cedex Bio HTCorrelation

Correlation• Good correlation between two

methods

• Small differences among clones(adjustment of factors possible)

• Reference method tends to resultin higher osmolality

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Osmolality Cedex Bio HTSummary

• More efficiency: Osmolality without extra sample, measurement, additional analyzer and time

• Good comparability with reference method (98% within ±10%, 76% within ±5%)

• Formula can be adjusted if needed (e.g. different matrix, or units in mg/L)

• Perfect for osmolality monitoring during fermentation process

Osmo = (2 x [Na+] + 1.3 x [Glc] + [Gln]) x 1.003

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Webinar agenda





SDC®

Sm@rtline Data Cockpit ® AGU 2018 ©

Sm@rtline Data Cockpit ®SDC

SDC Webinar June 28, 2018

Start

Sm@rtline Data Cockpit – a middleware for data integration Transfer of analytical data between devices and IT Systems like LIMS, ELN, MES.

Klaus BruchGeneral Manager

Jörg LehmannProduct Manager

Markus DückerDev. & Training

www.agu.de

SDC®


Sm@rtline Data Cockpit (SDC)

Main Goals

Goals:• Increase quality and reliability of laboratory data • Increase the throughput of your laboratory hardware• Reduction of the time for manually collecting data• Support to full fill the GMP requirements • Reduction off data integration costs

LIMS/ELN/MES

lab communication platform

SDC®


general situation in labs today

lost timeless accuracyand wasted resources

LIMS/MES/ELN/DCS

No data integration

youranalyzer

SDC®


each analyzer is integrated separately

GMP validation costs for each analyzerDevelopment costs for each analyzer type

LIMS/MES/ELN/DCS

Data Integration - direct

missing support for long barcodes missing entry fields like batch no missing support for multiple measurements missing electronic signature missing a common user management missing bidirectional communication

youranalyzer

SDC®


Smartline Data Cockpit ®21 CFR Part 11 compliance

LIMS/MES/ELN/DCS

validated validated validated

SDCmakes data integration easy

validated

Data integration with SDC

validated

Data Integration – SDC Middleware

network

Reduce validation costsReduce implementation timeKeep the upper systems simple one single

interface

Instrument layer

validated validated

support for long barcodes entry fields like batch no support for multiple measurements electronic signature (one/double) common user management

SDC is responsible for the interfaces!

youranalyzer

SDC®


Smartline Data Cockpit ® - ISInterface System

archivingof results

Smartline Data Cockpit ® - LSLaboratory System

select or create samples

archivingof results

Smartline Data Cockpit ®- LSLaboratory System

Possible registration of samples

transfer measurementsor control the analyzer

validation, repetition, approval, and

release of results

creation of measurements with the analyzer software

create measurementsBlack-Box

InterfaceInterface

LIMS/MES/ELN/DCS LIMS/MES/ELN/DCS

Templates

Templates

Comparison of the SDC operation modes (workflows)

SDC®


Cedex HiRes

Analyzer overview – Cedex HiRes

Cedex SoftwareCore

Cedex SoftwareUser Interface

SDC AIC Servicenetwork

• SDC can start and monitor the measuring process.− SDC considers samples with priority.− Interaction with the Cedex UI isn’t required.− SDC transfers additional manual measurements.

• SDC can start the maintenance functions.• Visualization of “runs left counter”.• Visualization of liquid status.• Scheduled execution of the database archive.

SDC®


Cedex Bio / Bio HT

Analyzer overview – Cedex HiRes

SDC AIC Servicenetwork

• SDC can send multiple measurement orders to the analyzer− Bio: Operator has to select the order in the display− Bio HT: no further operator input required− SDC gets results with detailed flag information

Bio HT v3.4:• SDC can start the maintenance functions.• Visualization of “runs left counter” and test status.• Processing of detailed calibration and liquid data

SDC®


Cell-Counters

Bio analysers

Blood gas analyser

Device overview

Cedex HiRes Vi-CELL™ XR

Cobas b 221

Cedex Bio HT Cedex Bio

Cobas b 123 BioProfile pHOx

BioProfile FLEX

RAPID Lab 248, 348, 348Ex, RAPID Point 500

Konelab 20, XT

Cedex Standard

ABL 800

coming soon

2950DBioProfile FLEX 2

coming soon

Logos, names and product images are protected and property-registered trademarks of their respective manufacturers.

SDC®


pH-Meter

Osmometer

Portavo 907Seven Excellence 3420 3310

3320 2020

auto 030 3000

Device overview

OsmoPro

coming soon

A2O

Osmometer


SDC®


Device overview

miscellaneous LAMBDA 25/35/45UV/Vis Spectrophotometer

miscellaneousClimet CI-153

DR 2800, DR 3900 photo meter Elecsys, cobas e 411

Palltronic FlowstarFilter integrity tester Octet® RED96

coming soon


SDC®


Device overview

ambr 15ambr 250

Process Control Advanced integration

Liquid Handler

coming soon

TECAN Sunrise

TECAN Fluent

Plate Reader

Automated Sampling

Microlab STAR Line

coming soon


SDC®


IT system overview

LIMS/ELN

Process ControlSystems

Historian

LIMSLIMSLIMSChemLMS

Generic SQLE-WorkbookAdvanced integration

MFCS/win

MES - SyncadeMES

Historian


SDC®


ambr data integration

ELISA reader

Osmometer

Roche Bio HT

• Full tracking of liquid handling operations• Creation of samplings and measurement orders with just a few mouse clicks.• Sending offline analytical results back to the ambr process control system• Prepared for auto working with automated sampling systems like

SDC

48 Batches

automatic

e.g. 142 measurements per sampling

Closing the feedback loop

48 samples in one sampling

Manual pipetting


SDC®


Thank you.

Contact

Klaus Bruch - graduate physicist [CEO]

Mobil: +49 (0) 151 151 487 13 Tel.: +49 (0) 214 868 60 13 Fax.: +49 (0) 214 868 60 19 E-Mail: [email protected]: www.agu.de

Jörg Lehmann – computer scientist[SDC product manager]

Tel.: +49 (0) 214 868 60 50 Fax.: +49 (0) 214 868 60 19 E-Mail: [email protected]: www.agu.de

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Disclaimer

Trademarks

For use in quality control/ manufacturing process only.

CEDEX is trademark of Roche. All other product names and trademarks are the property of their respective owners.

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Please contact your CustomBiotech representative

Visitcustombiotech.roche.com

Europe, Middle East, Africa, Latin AmericaPhone +49 621 759 8580Fax +49 621 759 [email protected]

United StatesPhone +1 800 428 5433, ext. 14649 (toll-free)Fax +1 317 521 [email protected]

CanadaPhone +1 450 686 7050Fax +1 450 686 [email protected]

JapanPhone +81 3 6634 1046Fax +81 3 5479 [email protected]

Asia PacificPhone +65 6371 6638Fax +65 6371 [email protected]

CustomBiotech from Roche

Thank you for your attention

Cedex Analyzers for Process Control in Bio-Processing · Common applications of monoclonal mouse...

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