LINDHOLT BIOSCIENCE

2 Contributors

SINGLE USE BIOREACTOR 1000L
Lindholt BioScience is looking to procure single use 1000L dual purpose Bioreactors that shall be designed to suite culture of Microbial / Mammalian organisms. The systems shall be installed in Multiproduct manufacturing area at customer site. Unit to be designed such that same Hardware and Software can be used for both mammalian as well as microbial culture. Software should have microbial and mammalian modes for operation. Bag holder to be designed to fit bags for mammalian and microbial culture. Also, other hardware like motor, MFC's, sparger to be designed for dual mode operation.
Technical requirements are prepared based on bioreactor to be procured for MPM-2 however number of equipment are more than required to be installed for MPM-2 as similar equipment shall be installed in other upcoming projects. Considering long delivery schedule equipment ordering is to be initiated now, later decision for installation to other areas shall be finalized.
Equipment Details
  • Working Volume: 1000L
  • Quantity: 8 Nos.
  • Location: Multi Product Manufacturing Facility at a customer site
PROCESS OVERVIEW
Typical process flows for various types of products that are intended to be manufactured in these Bioreactors is as follows:
Setup & Preparation
  • Setup the bioreactor and installation of the disposable bag
  • Calibration of Probes
  • Aseptically transfer of filtered media to the bioreactor bag
  • Setting parameters (RPM, Temperature, DO, pH)
Inoculation & Growth
  • Inoculation with culture
  • Sample analysis for OD
  • pH adjustment (with NaOH/Ammonia and Ortho phosphoric acid)
  • Feed solution addition and biomass build-up
Production Phase
  • De-repression solution addition (for yeast)
  • Induction solution addition (methanol for yeast, IPTG for E. coli)
  • Viral Infection (for mammalian cells)
Harvest
  • Collection of final product
  • System shutdown
Critical Process Parameters (CPP): pH, DO, Agitation speed, Temperature, weight
Critical Quality Attributes (CQA): Cell Concentration
CULTURE TYPES
Microbial:
Yeast Culture
The fermentation process of yeast culture would involve following steps:
  1. Setup the bioreactor and installation of the disposable bag
  1. Calibration of Probes
  1. Aseptically transfer of filtered media to the bioreactor bag from Vessel/bag using peristaltic pump
  1. Setting parameters i.e. RPM, Temperature, DO, pH
  1. Inoculation
  1. Sample analysis for OD, pH adjustment (with NaOH / Ammonia and Ortho phosphoric acid)
  1. Feed solution addition (Glycerol, salt mix, CMVT) and biomass build-up
  1. De-repression solution addition (Glycerol, salt mix, CMVT)
  1. Induction solution addition (methanol)
  1. Harvest
Microbial:
E. coli Culture
The fermentation process of E. coli culture would involve following steps:
  1. Setup the bioreactor and installation of the disposable bag
  1. Calibration of Probes
  1. Aseptically transfer of filtered media to the bioreactor bag from Vessel/bag using peristaltic pump
  1. Setting parameter i.e. RPM, Temperature, DO, pH
  1. Inoculation
  1. Sample analysis for OD, pH adjustment (with NaOH / Ammonia and Ortho phosphoric acid)
  1. Feed solution addition (glycerol, glucose) and biomass buildup
  1. Induction solution addition (IPTG)
  1. Harvest
Mammalian:
HEK293 Culture
The process of mammalian culture in the bioreactor would involve following steps:
  1. Setup the bioreactor and installation of the disposable bag
  1. Calibration of Probes
  1. Aseptically transfer of filtered media to the bioreactor bag from Vessel/bag using peristaltic pump
  1. Setting parameters i.e. RPM, Temperature, DO, pH
  1. Inoculation
  1. Sample analysis for OD, pH adjustment (with NaOH and CO2)
  1. Feed solution addition (glucose) and biomass buildup
  1. Viral Infection
  1. Harvest
TECHNICAL REQUIREMENTS
Operating/Functional Requirements
Single use bioreactor should have variable volume capacity (200L-1000L) and scalable H/D ratio. The system shall monitor and control gas flow rates, pH, DO, temperature, and agitation. Each MFC should have flexibility of interchangeability between Sparger1, Sparger 2 and Overlay.
Facility Requirements
Single Use Bioreactor shall be installed in a Grade B/C/D area with temperature 20-25°C and relative humidity 40-60%. Adequate space should be available to allow for operations, repair and maintenance activities.
System Requirements
All components, connections, and fittings shall be of sanitary design in accordance with ASME BPE Section SD-3. Agitation should be top driven/bottom driven with upward flow and downward flow capabilities.
Utility Requirements
Power supply: Single phase - 220V ±10% or Three phase - 415V ±10%. Non-condensing, oil and particle free Gases (Air, O2, CO2, N2) shall be available with required pressure ratings (2-4 Bar).
AUTOMATION REQUIREMENTS
Control System Features
  • List of alarms with provision to configure high/low alarm limits
  • E-stop mechanism to stop the batch immediately
  • Power failure & resume event recording
  • Protection of personnel, equipment & product in case of power failure
  • Data integrity during power failure events
  • Appropriate messaging during communication failure
  • Recipe continuation from point of interruption after power/communication resumes
Software Requirements
  • Different access levels as per 21CFR part 11 requirement
  • Configurable historian with auto data backup
  • Non-disableable audit trail recording all events/changes
  • Network environment compatibility
  • Time synchronization provision
  • Microsoft Windows based environment with English language support
  • Antivirus compatibility
  • Printer connectivity for real-time process data
The system should support recipe creation for operations including recipes for microbial culture, inoculation, de-repression mode, induction mode, mammalian culture, virus infection, and harvest. Cascade controls should be implemented for pH, DO, and de-repression processes.
Data Recording
Batch data including process parameters, alarms, and audit logs
Trending
Real-time visualization of all process actual values
Archiving
Storage of alarms, warning messages, and audit trail data
Reporting
Customizable reports with company details and process parameters
VALIDATION REQUIREMENTS
In order to verify system performance, the User shall witness the execution of the Factory Acceptance Test procedures. The Factory Acceptance Test Specification shall be submitted to the user for review and approval before 6 weeks of execution.
Factory Acceptance Test (FAT)
  • Mechanical component verification test
  • Electrical component verification test
  • Calibration verification for all measuring instruments
  • Bioreactor functionality testing
  • Access level and their rights verification
  • Utility and power failure test
  • Cultivation, Harvesting, Transfer sequence test
  • Printout verification / Audit Trail verification
Qualification Documentation
  • Design Qualification document from supplier
  • Software & hardware design specification document
  • FAT protocol, Project quality plan, traceability matrix
  • Installation Qualification documentation and execution
  • Operation Qualification documentation and execution
  • Vendor startup support and training
cGMP Compliance
  • WHO Guidelines TRS 979 (2013)
  • Drugs & Cosmetics Act, 1945, for schedule "M"
  • CE requirements and EU GMP, EN 285
  • US FDA 21 CFR part 11: "Electronic Records; Electronic Signatures"
  • EU GMP Volume 4, Annex 11 - Computerized systems
DESIGN SPECIFICATIONS
Bioreactor Controls and Accuracy
Agitation Specifications
Process Parameters
6
Mass Flow Controllers
Minimum quantity required for gas control
4
Peristaltic Pumps
Minimum quantity for media and solution additions
8
Total Units
Number of bioreactors to be procured
The Single Use Bioreactor design will follow industry, association and government codes. All aspects of the system will be compliant with current Good Manufacturing Practices (cGMP), the European Medicines Agency (EMA) guidelines, Good Automated Manufacturing Practices (GAMP), and ISO 9000 quality management standards.