BEC - 590 Interactive facility poster

Rituximab Biosimilar Production Facility

Interactive poster

Select a level to get started on the virtual facility tour- Shivani Sutrave Catherine Mathes Haroon Dar Arshad Ahmed

Floor 1

autoclave/ equipment sterlization room

Buffer and media prep room

Protein A chromatography

harvest

viral filtration

anion exchange chromatography

Viral Inactivation

bulk fill room

cation exchange chromatography

Lobby

200 L Reactors ( x 4 )

200 L Reactors ( x 6 )

waste management

This is the first floor of BTEC. As we enter through the doors we step into the Lobby which we have not touched, The drug substance moves from the Production suite into the Harvest, Protein A, Viral inactivation, CEX, AEX , viral filtration and bulk fill suites. All this movement is sequential and follows all the cGMP flows and regulations by the FDA.

Floor 1 - Personnel Flow

autoclave/ equipment sterlization room

harvest

Protein A chromatography

Buffer and media prep room

viral filtration

Viral Inactivation

bulk fill room

AEX

CEX

Lobby

200 L Reactors ( x 6 )

200 L Reactors ( x 4 )

waste management

Floor 1 - MATERIAl Flow

autoclave/ equipment sterlization room

harvest

Protein A chromatography

Buffer and media prep room

viral filtration

Viral Inactivation

bulk fill room

AEX

CEX

Lobby

200 L Reactors ( x 6 )

200 L Reactors ( x 4 )

waste management

250 ml shake flasks/ vial thaw

2 l shake flasks

buffer formulation /prep

Floor 2

5 l bioreactors

20 L bioreactors

cold storage room

Analytical Chromatography

Spectroscopy

servers

Imaging Suite

microscopy suite

pat

qc teaching lab

custodial

equipment sterlization

bioreactor instructional lab

autoclave

storage

conference

offices

training room

instructional lab

This is the second floor of the BTEC facility. Here we have the seed train expansion suites right across the stairs, buffer prep areas, training classrooms for our personnel, PAT section and offices for our employees.

Floor 2 - Personnel Flow

buffer formulation /prep

250 ml shake flasks/ vial thaw

2 l shake flasks

5 l bioreactors

20 L bioreactors

cold storage room

microscopy suite

Analytical Chromatography

Spectroscopy

servers

qc teaching lab

Imaging Suite

custodial

bioreactor instructional lab

equipment sterlization

autoclave

storage

conference

offices

training room

instructional lab

Floor 2 - material Flow

buffer formulation /prep

250 ml shake flasks/ vial thaw

2 l shake flasks

5 l bioreactors

20 L bioreactors

cold storage room

microscopy suite

Analytical Chromatography

Spectroscopy

servers

qc teaching lab

Imaging Suite

custodial

bioreactor instructional lab

equipment sterlization

autoclave

storage

conference

offices

training room

instructional lab

This is the second floor of the BTEC facility. Here we have the seed train expansion suites right across the stairs, buffer prep areas, training classrooms for our personnel, PAT section and offices for our employees.

Basement - Personnel Flow

Basement - Material Flow

Bulk Fill Room

This room will have an ISO 5 classified section as depicted. The eluate from the polishing chromatography steps is brought in here in bags, and processed in the ISO 5 sterile area and sent out to FujiFilm Diosynth for the final fill process. This room will have:

• Allegro Connect Bulk Fill System (1)
• ASI Aseptic Single-Use Filling System Model 4MP6000
• Watson Marlow 620 Di/R Pump
• Millipore M Air T air testing system (2)
• Climet CI-150t particle counter (2)
• Heraguard ECO Clean bench (2)

Viral Filtration

We will have the following main equipment here:

• Millipore ETU TFF skid (2)
• Millistak+ Pod Filter Holder (2)
• Millistak+ HC Pro Pods
• Parvovirus retentive filter

We will employ citric acid and Tris buffers and all the common utilities. Viral filtration, UF/DF will have classification of ISO 7. The AEX eluate is brought in here in carboys and we will utilize a parvovirus-retentive filter, which retains smaller viruses as well as larger viruses (20 nm+) while allowing mAbs to pass downstream.

Buffer Formulation/Prep

We will prepare all buffer/ media required throughout the processes in our facility in this room. This room will have the following main equipment:

• Feldmeier Media Prep tank V-4150 400 L
• Feldmeier Buffer prep tank V-4100 600 L
• Hyclone Mobius Mixer MX-1530 200 L
• Fisher refrigerator RF-1530
• Puffer Hubbard Freezer (3)
• Chemical fume hood
• Steril Gard II biosafety cabinet
• Watson Marlow 520s/R2 pump
• Sartocheck 4 integrity tester
• Ohaus scale (4)
• Fisher heat/stir plate (4)
• Hyclone stir plate

20 L Bioreactor Suite

This is our (n-1) production suite. We have the following equipment in here:

• Sartorius BIOSTAT Cplus 20L Bioreactor (10)
• Spare tubing + connectors
• Fisherbrand™ accumet™ Basic AB330 Conductivity/TDS/Salinity Benchtop Laboratory Meter
• Mettler Toledo FiveEasy Conductivity meter F30-Std-Kit
• BioPAT Trace Glucose/Lactate Measurement
• Computer with BioPAT MFCS SCADA software
• 50mL conical sampling tubes
• Biohazard waste bag and bin
• 4 degree C fridge

250 mL Shake Flasks/Vial Thaw

This is where the WCB will be thawed. We will have the following equipment in here:

• 250 ML ERLENMEYER CELL SHAKER FLASK ( x 30)
• Infors HT Mulitron Shaking Incubators (x 3)
• Pipette set
• Biohazard waste bag and bin
• Biological Safety Cabinets (2)

Harvest

The CCF from the Production step is transfered to this room in carboys. Here our desired secreted protein is removed from the cells and cell debris using a disc stack centrifuge, followed by depth and membrane filtration steps. This will have the following:

https://www.biopharminternational.com/view/harvest-and-recovery-monoclonal-antibodies-large-scale-mammalian-cell-culture

• Membrane filtration system
• Alfa Laval Disc stack centrifuge (1)
• Millistak+ HC Pro Pod Depth Filter C0SP
• MCE Membrane Filter, 0.22 um
• Carboys

Welcome to our rituximab biosimilar Facility

This is the new floor plan for the 2nd floor in this facility, navigate through different flows for 2nd floor with the icons below

Personnel Flow

Material Flow

Basement Personnel Flow

Basement Material Flow

Anion Exchange Chromatography Suite

The CEX pool is brought in carboys to this room & processed through AEX column. The AEX will operate in the flow through mode and it will be the second step in our polishing chromatography processes. This room will have:

• TOYOPEARL NH2-750F resin
• BPG Glass 300/500 column and skid (11)
• Sterile disposable bags
• Sterile tubing and gaskets

Spectroscopy

This room will have the following equipment:

• DXR3 Flex Raman Spectrometer
• Orbitrap Exploris™ 120 Mass Spectrometer
• Fisherbrand™ accumet™ Basic AB330 Conductivity/TDS/Salinity Benchtop Laboratory Meter
• Biohazard waste bag and bin

Cation Exchange Chromatography

The filtered VI pool is processed through an CEX column. The CEX operates in bind and elute mode, and we have 10 BPG Glass columns in this room processing in multiple cycles. This room contains the following equipment:

• TOYOPEARL Sulfate-650F resin
• BPG Glass 300/500 column and skid (11)
• Sterile disposable bags
• Sterile tubing and gaskets

Analytical Chromatography

This room will have the following equipment:

• Sartorius PATfix
• GE AKTA AVANT 150 CHROMATOGRAPHY SYSTEM
• GE AKTA AVANT 25 CHROMATOGRAPHY SYSTEM
• Sterile disposable bags
• Sterile tubing and gaskets
• Biohazard waste bag and bin

Welcome to our rituximab biosimilar Facility

This is the new floor plan for our facility.

Personnel Flow

Material Flow

Floor 2 Layout

Equipment Sterilization room

This room will have the following equipment:

• Steris GMP Autoclave
• Steris Decon Autoclave
• Branson Sonicator
• Lancer parts & glass washer

200 L Bioreactor Suite

Using ten Biostat D-DCU 200L twin Bioreactors with CIP and SIP, our production process in the 2 production suites culminates in Cell Culture Fluid transfer to the Harvest room.

• Biostat D-DCU 200L Bioreactors
• Hartel CIP Skid
• Denver Instruments pH/conductivity meter
• Thermo Scientific UV Spectrophotometer
• Nuaire biological safety cabinet
• New Brunswick Scientific incubator shaker
• YSI Biochemistry analyzer
• Scientific instruments vortex
• Biohazard waste bag and bin

Protein A chromatography

The HCCF is transferred here. ProA uses reversible and specific binding between immobilized protein A ligand and antibodies, it retains antibodies on the column, allowing elution in a purified and concentrated form. This room has the following equipment:

• TOYOPEARL AF-rProtein A HC-650 Resin
• Cytiva BPG Glass 300/500 Column and skid (11)
• Sterile disposable bags
• Sterile tubing and gaskets

Low pH Viral Inactivation

The proA eluate is subjected to viral inactivation which inactivates potential adventitious enveloped viruses by low pH treatment, incubation and the pH is readjusted and filtered. This room has the following equipment:

• Millipore ETU TFF skid
• Millistak+ Pod Filter holder
• Millistak+ HC Pro Pods

Welcome to our rituximab biosimilar Facility

This is the new floor plan for our facility.

Back to Floor 1

Material Flow

Floor 2 Layout

Welcome to our rituximab biosimilar Facility

This is the new floor plan for the 2nd floor in this facility, navigate through different flows for the 2nd floor with the icons below

Personnel Flow

Material Flow

Floor 1 Layout

Imaging Suite

This room will have the following:

• Cytation 5 system
• Thermo Scientific™ NanoDrop Lite Plus Spectrophotometer (2)
• Biohazard waste bag and bin

Microscopy suite

This room will have:

• Cytation 5 system
• Viranova Bioanalytics mini-TEM
• Micropipette set
• Biohazard waste bag and bin

Servers Room

The server room will encompass: DELL PowerEdge R7525 Rack Server (x15) 25U Server rack Uninterrupted Power Supply

5 L Bioreactors

This room employs the following equipment:

• Sartorius BIOSTAT Cplus 5L Benchtop Reactors (15)
• Biological safety cabinet
• Micropipette set
• 2mL sampling syringes
• 4-degree C fridge
• Biohazard waste bag and bin

2 L Shake Flasks

We will transfer the cells to this room for scale-up. These are the main equipment in this room:

• 2L Erlenmeyer Flask w/ Vent Cap, Polycarbonate, Sterile (x 24)
• Infors HT Mulitron Shaking Incubators (x 3)
• Pipette set
• Biohazard waste bag and bin
• Biological Safety Cabinets (2)

Welcome to our rituximab biosimilar Facility

This is the new floor plan for the Basement in this facility, navigate through different flows for the basement with the icons below

Basement Personnel Flow

Basement Material Flow

Floor 1 Layout

Floor 2 Layout

Cold Storage room

Any extra cold storage requirements will be met here. This room will have the following equipment:

• Fisher refrigerator RF-1530
• Puffer Hubbard Freezer (x 2)
• Steril Gard II biosafety cabinet

Buffer / Media Prep room

Our buffer room formulates diverse buffers crucial for upstream processes, regulating pH, optimizing cell growth, and preserving protein functionality during primary recovery in preparation for purification. It will have the following equipment:

• Feldmeier Media Prep tank V-4150 400 L
• Feldmeier Buffer prep tank V-4100 600 L
• Hyclone Mobius Mixer MX-1530 200 L
• Fisher refrigerator RF-1530
• Puffer Hubbard Freezer (3)
• Chemical fume hood
• Steril Gard II biosafety cabinet
• Watson Marlow 520s/R2 pump
• Sartocheck 4 integrity tester
• Ohaus scale (4)
• Fisher heat/stir plate (4)
• Hyclone stir plate

Welcome to our rituximab biosimilar Facility

This is the new floor plan for the Basement in this facility, navigate through different flows for the basement with the icons below

Basement Personnel Flow

Basement Material Flow

Floor 1 Layout

Floor 2 Layout