The CuBiAn XC – A universal tool for your bioprocess analysis Photometric, automated, XC: Extra Compact photometrisch, automatisiert, Gelöste Substanzen: Substrate, Metabolite, Aminosäuren, Protein CellD 20bis, rue du Chapitre F-30150 ROQUEMAURE Tel : +33 (0)4 66 82 82 60 Fax : +33 (0)4 66 90 21 10 Email : contact-info@celld.com www.celld.com Alexander Hähnel Sandra Koptik

The CuBiAn XC presentation Sample & Reagent Handling Accuracy and precision The assay portfolio The IgG assay principle The validation package Comparison membrane - photometry Automation capability of CuBiAn® XC

Incubation Reaction Unit Overview over the analyser Sample & Reagent Handling Robotic Cuvette Arm Cuvette Platform Reagent and Sample Pipette Unit Mixing Unit Photometric Detector Unit Reagent and Sample Container Unit Incubation Reaction Unit (below cover)

Sample & Reagent Handling Simple, automated handling or reagents and samples Sample & Reagent Handling Most reagents ready-to-use Reagents and samples kept refrigerated on board Reagents and samples optionally identified by barcode reader (type, expiry date, and consumption) Reagents single assays obtainable separately for maximum flexibility Each reagent bottle allows for 96 tests Automatic dilution steps for samples programmable Low sample volume required: 2µL - 35µL per test ≤ 200µL minimum volume The Reagent and Sample Container Unit has space for 20 assay bottles and 10 samples

Sample & Reagent Handling The measurement workflow Sample & Reagent Handling Samples and reagents automatically pipetted into disposable cuvettes in the Incubation Reaction Unit (IRU) Each reaction is mixed thoroughly Reactions constantly incubated at 37 °C The cuvettes will be uploaded, checked before use and discarded by the automated robot arm Samples can be run in replicates (2-40) Rapid delivery of results Up to 90 results/hour possible (360 with ISE unit) Assays measured every 40 seconds Each reaction takes exactly 10 minutes

Accuracy and precision Liquid handling Accuracy and precision The automated pipetting system for reagents and samples guarantees for a high reproducibility and accuracy The reagent and sample syringes work with a cv of less than 1%, and 2% respectively The accuracy of pipetting can be checked at any time by the user via (SPT+ RPT (Sample and Reagent Precision Tests; part of the CuBiAn® SST concept)

Accuracy and precision Calibrator and Control liquids Accuracy and precision Two multi- (universal) calibrators create calibration curves for the majority of the assays (Lactate, Glucose, Ammonia, Glutamate, Inorganic Phosphate, Iron, Mg, Cholesterol,...) Separate Calibrators for Glutamine and IgGs Two or three levels of controls (low, medium, high) available to check the system’s precision of the specific assay

Accuracy and precision Calibration graphs Accuracy and precision

Accuracy and precision Control status documented in quality graphs (Levey-Jennings plots) Accuracy and precision

Accuracy and precision Customizable warning scenarios Accuracy and precision

The assay portfolio Extract of current assays and ranges (hs) IgG 10.0 – 500 mg/L (hc) IgG 2.8 – 27.55 g/ L Total Protein 0.04 – 2.25 g / L Glucose 0.09 - 6.3 g/L Lactate 0.05 – 8.0 g/L Ammonium 0.05 – 9.0 mM Glutamine 0.06 – 10 mM Glutamate 0.12 – 10 mM Inorganic Phosphor 0.13 – 10.32 mM LDH (L->P; P->L) 13 – 2800 U/L Na+ 80-180 mM K+ 1.5 - 10 mM CL- 60-140 mM Reagents ISE unit Reruns and pre-dilutions allow to enlarge the measurement ranges displayed above

Recently launched, in final tests, under development The assay portfolio Acetic Acid (launched; 0.05 – 1.25 g/L) Magnesium (launched; 0.2 -3.27 mMol/L) Cholesterol (launched; 0.87 – 16.6 mMol/L) Iron (launched; 2 – 198 µMol/L) Glycerol (final tests; 0.016 -2.5 mMol/L) Galactose (under development: 0.05 – 7 g/L)

The IgG assay principle Principle of immuno turbidimetry The IgG assay principle Model of a human IgG Target of the anti-serum in the hs IgG assay

The IgG assay principle Principle of immuno turbidimetry The IgG assay principle

The IgG assay principle Parameter settings customizable – Instrument Factor The IgG assay principle

The IgG assay principle Comparison measurements against HPLC The IgG assay principle 10 / 23

The Validation Package Validation documents are available (Verification documents including IQ, OQ); PQ on demand Experienced validation consulting and support service by OPTOCELL technology Documents developed in close cooperation with customers

Photometric vs. Membrane based analysers Comparison to membrane based analysers Photometric vs. Membrane based analysers Membrane based (electrochemical) systems CuBiAn® photometric & immuno-turbidimetric systems Expandability Limited Nearly unlimited and customizable Necessary Frequency of Calibration Very high Once per lot of reagents Reproducibility of Results Low High Immuno-turbidimetry Not possible Implemented On-line capability Limited due to chemical sterilization BayChromat® CellCount + CuBiAn®XC with steam sterilized sample valve Rate analysis

CuBiAn® photometric and immunoturbidimetric Systems Comparison to membrane based analysers Photometric vs. Membrane based analysers CuBiAn® photometric and immunoturbidimetric Systems Membrane based electrochemical (biosensors) systems

Photometric vs. Membrane based analysers Comparison to membrane based analysers Photometric vs. Membrane based analysers

Automation capability The automation capability of the CuBiAn XC Automation capability The CuBiAn® XC can be completely integrated into a fully automated bioreactor environment without any technical hardware changes The remote access software (developed together with Bayer Technology Services, Germany) can be easily upgraded to an CuBiAn® XC The steam-sterilizable automatic sample drawing system of the BayChromat ® Cell Count has shown its high stability in several contamination-free cultivations, lasting over more than 180 days The BayChromat ® control software allows for the integration of additional analysing devices

Automation capability Comparison to membrane based analysers Automation capability The BayChromat® Cell Count by: Bayer Technology Services, Leverkusen, Germany The STAT port

Outlook – CuBiAn Family Throughput: 270 tests/hour (max. 404 tests/hour with ISE High precision pipettor und reagent pipettor Environment-friendly semi-disposable resin cuvettes Refrigerated Reagent Container Unit Optional built-in ISE module available Optional built-in degaser module available