LINDHOLT BIOSCIENCE

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.