1 | U p s t r e a m B i o P r o c e s s i n g Q u e s t i o n n a i r e

Upstream BioProcessing Questionnaire

Please answer the following questions as completely as possible.

I. Customer Information

Contact Person

Designation

Department

Company Name

Contact Number

Email Address

II. General Details

1. Target Product □ Secreted Protein □ Non-secreted Protein □ Cell bank □ Monoclonal Antibody □ Secreted Virus □ Non-secreted Virus □ Autologous Cell Therapy (Please answer Cell

Therapy Questionnaire) □ Allogenic Cell therapy (Please answer Cell

Therapy Questionnaire) □ Others:

2. Cell Type □ Adherent cell (Proceed to Adherent Cell Questionnaire)

□ Suspension cell (Proceed to Suspension Cell Questionnaire)

□ Microbials (Proceed to Suspension Cell Questionnaire)

□ Stem Cell

Please fill out questionnaire respectively depends on EACH application.

(Different cell line, different application has to be filled in separate questionnaire).

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Adherent Cells Questionnaire

I. Experiment Details

a. General Details

1. Cell Line □ CHO □ MDCK □ Vero □ HEK 293 □ Hybridoma □ Sf 9 □ Others:

2. Any special features or peculiarities of the cell line or culture methods

3. Intended Use □ Human Use □ Animal Use

4. Target Product □ Secreted Protein □ Non-secreted Protein □ Cell bank □ Monoclonal Antibody □ Secreted Virus □ Non-secreted Virus □ Others:

5. Current Culture System □ T-flask

____________ cm2 x ____________ Pcs

□ Petri dish

Diameter _________ mm x ____________ Pcs

□ Roller bottle

____________ cm2 x ____________ Btls

□ Spinner flask

____________ ml x ____________ Btls

Carriers: ________________________

□ Cell factory (multi-layer) ____________ cm2 x ____________ Pcs (Total surface area)

□ Stirred-tank Bioreactor

____________ ml x ____________ vessel

Carriers: ________________________

□ Others: Total Volume Capacity: _________________ L

6. If carriers are used, please specify type and amount of carrier.

□ Microbeads, Specify: □ Fibers, Specify: □ Others, Specify:

Amount of carrier:

7. Working Volume Capacity ________________________ mL

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8. Medium change frequency for current system

□ 24 hours (1 day) □ 48 hours (2 days) □ 72 hours (3 days) □ Other : ____________ hours (_____ days)

Media volume per change: _____ ml

9. Cell Culture Condition □ Media:

□ Serum:

□ Temp.:

10. Concentration of additives □ Sodium bicarbonate:

□ Hepes buffer:

□ Others:

11. Glucose Concentration in initial culture medium

________________________ g/L

12. Cell Harvesting (Cell Dissociation) Required

□ Yes □ No

13. Cell Harvest (Cell Dissociation) Method

□ Trypsin □ Enzymatic Dissociation Reagents

(Specify: ________________________)

□ Non-Enzymatic Dissociation Reagents

(Specify: ________________________)

□ Others

14. Cell Quantification (Cell Counting)

□ Manual counting □ Auto-counter □ Nuclei counting □ Others:

15. Access to a bio-analyzer for measuring glucose, lactate, glutamine, etc

□ Yes □ No

16. System preference □ Prefer Single-Use □ Prefer Multiple-Use □ No Preference

17. Expected annual dose (product quantity)

18. Expected Total Cell number from current system (for the application that harvest cells)

________________________ cells

19. Do you have scale up plan □ Yes

Planned scale up year

□ No

20. Expected Scale when scaled-up (Cell number, Doses etc)

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b. Protein Production

1. Culture period prior to harvesting

□ 24 hours (1 day) □ 48 hours (2 days) □ 72 hours (3 days) □ 96 hours (4 days) □ Other: _____ hours ( _____ days)

2. Protein extraction method □ Cell Harvest □ Medium Harvest □ Freeze/Thaw □ Others:

3. For medium harvest, what is the harvest method?

□ Single Harvest □ Multiple Harvest

Interval time ____________ hrs for

____________ days

c. Virus Production

4. Virus Strain

5. Please describe the Virus Strain? (ds,ssDNA, ds,+/- ssRNA, enveloped, non-enveloped, temperature sensitivity, etc.)

6. Does cells stable during post infection?

□ Yes, grow and attach same as before infection □ A little less stable than before infection □ No, cells tend to detach post infection period

____________ hours

7. What is the temperature of virus be active

____________ oC - ____________ oC

8. Does virus MOI sensitive of temperature difference?

□ Yes □ No □ Not sure □ Others:

9. Cell density prior to infection in current culture system

10. Do cells propagate after virus infection?

□ Yes Fold increase post infection: ____________

□ No □ Not sure

11. Multiplicity of Infection (MOI)

12. Virus titer in current culture system

________________________ dose/ml

13. Period of time for cell lysis to occur after infection in current culture system

________________________ hours

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14. Culture condition post infection

□ Media:

□ Serum:

□ Temperature:

□ Others

15. Glucose Concentration in culture medium

________________________ g/L

16. Best phase for infection □ Seed cells with virus □ Right after seeding □ Exponential phase □ Plateau phase □ Not sure (____________ hours after cell

culture)

17. Best time to harvest the virus

18. Is there CPE (cytopathic effect) after infection? When?

□ Yes _____ hours / days later □ No

19. What kind of CPE(cyto-pathic effect) is formed

□ Monolayer Destruction □ Swelling □ Clumping □ Vacuolization □ Inclusion bodies □ Others:

20. Cell lysis post infection □ Yes _____ hours / days later □ No

21. Number of harvests that could be done during post-infection period

□ Single Harvest □ Multi-harvest

_____ times with ______ interval hours

Each time harvest volume: ____________ L

□ Continuous Harvest

_____ days with total volume __________ L

22. Do cells keep propagation after virus infection?

□ No □ Yes, Indicate fold increase post infection:

d. CelCradle™ System

1. Will seeding of 1 x 108 cells be difficult?

□ Yes □ No

If yes, how many cells do you plan to seed?

2. Will the CO2 incubator be exclusively used for the CelCradle™ sytem?

□ Yes □ No

Can you adjust the CO₂ concentration?

□ Yes □ No

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Fermentation/Suspension Cells

I. Experimental Details

a. General Details

1. Application □ Microbial Culture □ Suspension Cell Culture □ Adherent Cell Culture

2. Cells Culture □ Bacteria □ Yeast □ Fungi □ Cell Line: □ Other:

3. For adherent cells, specify type and amount of carrier used

□ Microbeads, Specify: □ Fibers, Specify: □ Others, Specify:

4. Amount of carriers per batch

5. Target Product □ Secreted Protein □ Non-Secreted Protein □ Fermentation □ Cell Banking □ Secreted Virus □ Non-Secreted Virus □ Others:

6. Current Culture System □ Spinner Flask □ Stirred Tank Bioreactor □ Other:

7. Current Culture Seeding Volume

8. Current Culture working volume

________________________ L

9. Current Culture Total volume

________________________ L

10. Current Culture Yield

11. Current Process Mode □ Batch □ Fed-Batch □ Continuous □ Other:

12. Current Culture Period (from seeding to harvesting)

□ 24 hours (1 day) □ 48 hours (2 days) □ 72 hours (3 days) □ Others: ____________hours (_______ days)

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13. Culture condition for cell growth

□ Media: □ Serum: □ Temp.:

14. Concentration of additives □ Sodium bicarbonate: □ Hepes buffer: □ Others:

15. Glucose Concentration ________________________ g/L

16. Cooling system required? □ Yes □ No

17. Temperature sensitive? □ Yes □ No

18. pH sensitive? □ Yes □ No

19. Shear Stress tolerance □ High Sensitivity □ Medium Sensitivity □ High tolerance to shear stress

20. Preferred Impeller type and impeller distance to vessel

□ No preferrance □ Rushton Turbine □ Marine Blade □ Pitched Blade □ Spin Filter □ Others

Distance to vessel

21. Agitator required? □ Yes □ No □ Not sure

22. Do you use anti-foam reagent?

□ Yes (Specify: ________________________)

□ No

23. Broth Rheology □ Newtonian □ Non-newtonian □ Not sure

24. Maximum OTR (oxygen transfer rate)

25. DO (dissolved oxygen) requirement

26. Maximum Broth Viscosity

27. Final broth volume

28. Regulation / BSL Level / Containment Level

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b. Process Control

1. Reactor Size Minimum working volume: ____________ L Maximum working volume: ____________ L

2. Agitation Speed Range: ____ rpm to ____ rpm

3. Measurements Required □ Temperature ____ oC to ____ oC

□ pH ____ to ____

□ DO ____ % to ____ %

□ Redox ____ mV to ____ mV

□ Turbidity

□ Foaming

□ Level

□ pCO2

□ O2/CO2 in Exhaust Gas

□ Others

4. Temperature Control Range ____________ to ____________ oC Control:

□ Double wall vessel □ Heating Jacket □ Heating Pad □ Heating/Cooling Pad □ Other:

5. pH Control Range ____________ to ____________ Control:

□ Addition of Base □ Addition of Acid □ Addition of CO2 □ Others:

6. Dissolved Oxygen Control Range ____________ to ____________ % Control:

□ Impeller Speed □ Addition of O2 □ Addition of N2 □ Gas Flow Rate □ Others:

7. Foaming □ High □ Low □ Not yet determined

8. Applied Gases for Aeration □ Air □ Air + O2 □ Air + O2 + N2 □ Air + O2 + N2 + CO2 □ Others

Mixing System: □ Yes □ No

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9. Airflow Range: ____________ to ____________ vvm

Control:

□ Regulator (manual) □ Mass Flow Controller □ Others:

10. Aeration Delivery □ Overlay □ Sparger, Type:

o Ring Sparger o Microsparger o Others:

□ Both

11. Pressure Control Requirements

12. Pressure Control System □ Manual Control □ Automatic Control □ Others:

13. Overpressure Control: □ Release Valve □ Rupture Disk □ Others:

14. Hydrostatic Pressure

15. Other Special Requirements

c. Reactor Requirements

1. Vessel Material □ Borosilicate Glass □ SS 316L □ Others:

2. Pressure vessel standard

3. Vessel Geometry □ 1:2 □ 1:3 □ Other:

4. Internal coils □ Yes □ No □ Not sure

5. Maximum coolant flow temperature

________________________ oC

6. Minimum fluid temperature ________________________ oC

7. Compressor max press and max flow

Max press: Max flow:

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8. Seeding □ Needleless Seeding Port □ Needle Injection Inoculation Port □ Others:

9. Pressure Control System □ Manual Control □ Automatic Control

10. Sterilization Temperature: ____ oC to ____ oC Period:

11. Fluid Addition Volume: Number of Ports:

□ 0 □ 1 □ 2 □ 3 □ 4 □ Others:

12. Number of Sampling Port □ 0 □ 1 □ Others:

13. Air Filter Housing Filter Size: □ 0.2µm □ Others:

Integrity Test Port: □ Yes □ No

14. Exhaust Filter Housing Filter Size: □ 0.2µm □ Others:

Integrity Test Port: □ Yes □ No

15. Impeller Motor □ Top driven (standard) □ Bottom driven

Speed: ____________ to ____________ rpm

16. CIP requirement □ Yes (__________________________________)

□ No

17. Space dimension (end user)

□ Length: ____________

□ Width: ____________

□ Height: ____________

d, Control Requirements (Esco VacciXcell standard is Siemens HMI / PLC TP 1500)

1. Siemens SCADA □ Yes □ No

State Brand used:

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2. Siemens PCS 7 □ Yes □ No

State Brand used:

3. Warehouse management □ Yes State Brand used:

□ No

4. Historian Archive □ Yes □ No

5. Compliance 21 CFR part 11 / Gamp 5

□ Yes □ No

6. E-batch records □ Yes □ No

7. Digital Signatures □ Yes □ No

8. CSV required □ Yes □ No

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Cell Therapy

I. Experiment Details

a. General

1. Target □ Autologous Cell Therapy □ Allogeneic Cell Therapy □ Secreted Protein

Specify: ________________________ □ Research □ Others:

2. Cell Source □ Bone Marrow □ Adipose-derived □ iPS □ Embryo □ Placenta □ Umbilical □ Dermal fibroblast □ Others:

3. Any special features or peculiarities of the cell line or culture methods

b. Current Culture

1. System Used □ T-flask ____________ cm2 x ____________ Pcs

□ Petri dish Diameter _________ mm x ____________ Pcs

□ Roller bottle ____________ cm2 x ____________ Btls

□ Cell Factory (multi-layer) ______________ cm2 x ____________ Pcs (Total Surface Area)

□ Spinner flask

____________ ml x ____________ Btls

Carriers: ________________________

□ Stirred-tank Bioreactor

____________ ml x ____________ Vessel

Carriers: ________________________

□ Others: Total Volume Capacity: ____________ L

2. Working Volume Capacity ________________________ mL

3. If carriers are used, please specify type and amount of carriers

Type: □ Microbeads □ Fibers □ Others

Amount of carriers:

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4. Current culture method □ Batch □ Fed-batch □ Recirculation □ Continuous □ Others: ________________________

5. Medium change frequency for current system

□ Do not change media until harvesting □ 24 hours (1 day) □ 48 hours (2 days) □ 72 hours (3 days) □ Other: ____________ hours

Media volume per change: ____________ mL

6. Culture Condition □ Media: □ Serum: □ Temp.: □ CO2 conc.: □ Initial Glucose Conc.: ____________ g/L □ Others:

7. Culture Medium □ Serum-containing □ Serum-free □ Xeno-free □ Protein-free □ Animal Component -free

8. Number of days of culture (from seeding to harvesting)

□ 24 hours (1 day) □ 48 hours (2 days) □ 72 hours (3 days) □ 96 hours (4 days) □ Others: ________________________

9. Cell Harvest (Cell Dissociation)

□ Trypsin □ Enzymatic Dissociation reagents

(Specify: ________________________) □ Non-Enzymatic Dissociation reagents

(Specify: ________________________) □ Others: □ No harvesting required

10. Cell seeding total number

11. Cell harvest total number per culture device

12. Number of culture device used per batch (patient or treatment)

13. Cell Quantification (cell counting)

□ Manual Counting □ Auto-counter □ Nuclei counting □ Others

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c. Process Plan

1. System Preference □ Prefer single-use □ Prefer multiple-use □ Prefer hybrid □ No preference

Remarks:

2. Do you have scale up plan? □ Yes Pipeline

□ Not in this moment

3. Expected harvesting cell number when sale up

________________________ cells □ 109 □ 1010 □ 1011 □ 1012 □ 1013 □ Other: