Plant Design and Manufacturing Principles in DNA Vaccine Production


Bachelor Thesis, 2017

37 Pages, Grade: 80.0


Excerpt

Contents

Abstract

1.0 Introduction
1.1 Aims/objective

2.0 Detail of the process
2.1 Process description
2.2 Raw material required
2.3 Equipment required
2.4 Staff Involved

3.0 Design of Manufacturing facility
3.1 Plant location
3.2 Plant layout
3.21 Layout of the Plant site appendix 1
3.22 Layout of the Production House appendix 2
3.3 Manufacturing Flow.
3.4 Room Specification data sheets appendix 3

4.0 Plant Services, Systems and Utilities
4.1 Heating, ventilation and air-conditioning (HVAC) system
4.2 Water
4.3 Clean steam.
4.4 Heat and power
4.5 Cleaning Systems
4.51 Clean in Plane (CIP)
4.52 Sterilise in Place (SIP)
4.6 Cleanrooms
4.7 Process control & instrumentation Instrumentation

5.0 Process Validation and cGMP
5.1 Process Validation
6.0 Gannt Chart
7.0 Costing
8.0 Conclusion
9.0 References

Appendix
Appendix 1 Layout of the Plant site
Layout of the Production House Appendix 2
Gantt Chart Appendix 4
Costing Appendix 5

Abstract

The demand for DNA vaccines in large quantities at high purity for gene therapy is on the increase. As it helps to stimulate antibodies production in human and provide immune protection against many diseases such as cancer, malaria, HIV and other diseases (Laere et al., 2016) and have potential advantages over conventional vaccines.

Therefore, this report covers a detail design and cost (to an accuracy of +/- 20%) for a new manufacturing facility to produce DNA vaccines to be built on a greenfield site. Applied current good manufacturing practice (cGMP) and complied with all the regulatory guidelines set up by various agencies.

A process to produce commercial pDNA covers: Fermentation, recovery and product purification. The layout of the manufacturing facilities has been designed in a way to allow a good waste, raw material and personnel flow to minimise the risk and contamination. All of these are an example of current good manufacturing practices which are vital for the production of the therapeutic product (DNA vaccines).

Plant services, system and utilities has been designed to meet the requirement for the manufacturing facility which includes Water for CIP to clean equipment and media preparation, Clean steam for SIP to sterilise the equipment, Heating, ventilation and air conditioning (HVAC), water, Compressed air, Effluent treatment, Nitrogen, USP, CIP system, refrigeration and more. (Cole, 1998).

The facility will take up to 2.6 years to complete and qualify (GSK, 2017). The project schedule will depend on various activities these must be completes to produce DNA vaccine facility, ready for start-up. These includes Front end design, Detailed design, Procurement of equipment, construction, recruitment of staff, commissioning of facility and validation of process. (Adkin, 1998)

Finally, the costs of the pharmaceutical plant are a serious consideration when planning whether a plant will be commissioned or not. The economic evaluation for the DNA Vaccine Plant suggests that the plant will be at significant profit with a payback time of less than 2 years.

1.0 Introduction

DNA Vaccines helps to stimulate antibodies production in human and provide immune protection against many diseases such as cancer, malaria, HIV and other diseases (Laere et al., 2016) and have potential advantages over conventional vaccines. The demand for DNA vaccines in large quantities at high purity for gene therapy is on the increase. However, inaccessibility of DNA vaccines for the treatment of diseases can cause several problems. Therefore, to meet the rising demand for DNA vaccines, there is a need for designing & manufacturing facilities to produce DNA vaccines. In order to fulfil the need for the production of DNA vaccines the manufacturing facility such as design, operation and layout must comply with the regulatory requirements stipulated that the production of DNA vaccine must be performed under current good manufacturing practices (cGMP) and good design practices (GDP). Also, comply with the guidance regarding the specification, quality testing and manufacturing standard of DNA products by regulatory agencies such as the US Food and Drug Administration (FDA) and the European Agency for the Evaluation of Medicinal Products (EMEA) and also regulation stipulated by the country where the facility is being constructed. (Przybylowski et al., 2017)

According to the EU guide, GMP is the part of Quality Assurance (QA) which make sure that products are consistently produced and controlled to the quality standards appropriate to their intended use and as required by marketing authorisation or product specification (Institution of Chemical Engineers (Great Britain), 2003).

When designing a new drug production manufacturing facility, the following consideration needs to be made (GMP requirements). To ensure the safety, identity, purity and potency of the manufactured product (Przybylowski et al., 2017). Not placing the patient at risk due to insufficient safety or quality.

- Establishment of effective QA system
- Control of the process
- Personnel that are competent, appropriately qualified and trained.
- Facilities – suitable for operation, layout, design and operation reduces the risk of errors. Cleaning in place (CIP)
- Premises and equipment that are critical for product quality are validated (qualified).
- Standard operating procedure to Prevent contamination from any sources such as environment, equipment and premises.
- Environmental impact assessment (EIA)
- Documentation and audit of all aspects of the process.

1.1 Aims/objective

- Propose concept designs and costs (to an accuracy of +/- 20%) for a new manufacturing facility to produce DNA vaccines to be built on a greenfield site. Apply current good manufacturing practice (cGMP) and comply with all the regulatory guidelines set up by various agencies.
- Schedule for the project / timeline of the project.
- The facility design will include – Design of manufacturing facility, manufacturing flow, plant services system and utilities, Process validation & cGMP and costing.

2.0 Detail of the process

A process of commercial pDNA manufacturing includes: Fermentation, recovery and product purification. Below figure 1 shows the unit operation involved in the manufacturing process.

2.1 Process description

Abbildung in dieser Leseprobe nicht enthalten

Figure 1 : the process flow of the production of therapeutic plasmid purified bulk products. Adkin, A. (1998). Design of a multi-use DNA vaccine production facility. Project report submitted for partial fulfilment for the degree of Master of Science in Biochemical Engineering., [online] 1(UCL). Available at: https://www.researchgate.net/publication/260581752_Design_of_a_multi-use_DNA_vaccine_production_facility [Accessed 5 Mar. 2017].

Batch Fermentation – Fermentation process is the starting point for the production of pDNA in the bio-reactor, and the aim is to maximise pDNA productivity per unit cell mass. This takes place under the optimum condition to maximise the cell growth. (Adkin, 1998)

Received raw material is placed in quarantine unit until clearance from QA. Production culture stains within the facility are maintained as MCB and WCB’s. At the start, bacteria from a WCB’s vial is revived and grown in a propagation area up to 2 cultures. Sterilised prepared media is feed into the fermenter. Two seed vessel of volume 15L and 150L are used to produce inoculum for the 1000L cell culture vessel. Operated in the fed-batch mode to provide product for successive downstream processing with operating temperature of 37oC optimum for bacteria. Upon the establishment of maximum cell growth. The broth is cooled to 40C and pumped to centrifuge at a flowrate of 1000L/h, as this allows effective separation of cellular material. Concentrated broth contains pDNA and some other cell organelles; this is discharged in the alkaline lysis vessel. (Adkin, 1998)

Alkaline Lysis – three step process. 1) concentrated broth is suspended in the buffer solution for alkaline lysis - low viscosity. 2) The cell suspension is lysed by mixing with NaOH/SDS – solution changes highly viscous. 3) 3M potassium acetate neutralisation causing the precipitation of impurities such as chromosomal DNA whilst minimising the loss of pDNA yield, the solution becoming Newtonian again. Bag Filtration – Disposable non-leaching nylon bags doesn’t contaminate the product, fully scaleable. This partially removes the solid precipitate from the lysate before further product purification.

Anion Exchange chromatography – separation occurs due to electrostatic attraction between the solute and charged dense clusters. Anion exchange chromatography clears all the cationic proteins and lipopolysaccharides and retains all forms of pDNA, the residual cDNA, RNA and any anionic proteins. (Nigel, 2017)

Gel filtration – The removal of small RNA, aggregates or clipped products by Size Exclusion Chromatography; a non-adsorptive chromatography process that separates cluster that differs in size from pDNA in desalting solution. (Nigel, 2017)

Endotoxin Removal – the produced product should be free from all sort of contaminants for the purposes of therapeutic. Therefore, the final step includes the removal of anionic endotoxin by a particular adsorptive chromatographic process. (Nigel, 2017)

Finally, formulating the product by putting stabiliser, correct salt concentration and sterile filtration & lyophilisation.

2.2 Raw material required

The following Raw materials are needed for the production of DNA vaccines. All the raw materials entering the process should be controlled and tested to make sure the manufacturing process meet the standard requirements. The most important Raw material is plasmid DNA. Therefore, it should be fully characterised. For Therapeutic pDNA E. coli is the most common host. (Przybylowski et al., 2017)

Raw materials required – pInG human tyrosinase (pING-HT) plasmids, E.coli, Nutrient - Glucose, antifoam,water, liquid nitrogen, ammonia hydroxide, Ethylenediaminetetra acetic acid diammonium salt (EDTA), Hydrochloric Acid, Magnesium Sulphate, Polypropylene Glycol (PPG), Potassium Acetate, Potassium Phosphate, Sodium chloride, Sodium Hydroxide and Tris Hydroxymethyl methylamine (TRIS) buffer. (Adkin, 1998)

2.3 Equipment required

The following table shows the manufacturing equipment required for the process, these should be designed, located and maintained to suit its intended purpose and these should not affect the quality of the product.

Abbildung in dieser Leseprobe nicht enthalten

Table1: Equipment used in the process (Adkin, 1998)

2.4 Staff Involved

The correct manufacturing of the product (DNA Vaccines) depends upon people/staff. Therefore, qualified personnel must carry out all the task which are the responsibility of the manufacturer. The manufacturer should have enough staff with essential qualification and experience, and they should be aware of cGMP and receive appropriate training. (Inspection and Healthcare, 2002)

All the staff members whose activate could affect the quality of the product should be provided with adequate and continuing training such as technical, maintenance and cleaning staff.

Key personnel include Head of Production and Head of Quality Control (QC). Other personnel include Process Engineer, Technician, Maintenance, Operative, Assistance and cleaning. (Inspection and Healthcare, 2002)

3.0 Design of Manufacturing facility

The design of the new manufacturing site for (DNA Vaccine) will be laid out for primary production, secondary production, research and development, warehousing and distribution/administration and head office activities. (ICheme, 2003)

3.1 Plant location

The location of the drug production manufacturing facility is crucial to its profitability as it can be affected by raw material supply, climate, transportation, utilities and services, etc.

Site selection

The DNA vaccine plant will be located in southern England, Slough. This is chosen because the site is relatively near to greater London which can create huge advantages which include:

- Transport – the site is located by M25, which will allow cost effective transportation. The site is also located near to the Heathrow airport; this will allow cost-effective exports (Products) and imports (Raw materials) while timesaving.
- Labour - The site is relatively near to greater London which has high-density population. Therefore, there will be plentiful labours for the plant, and there is good transport link.
- Site cost – as the land is outside the vicinity of London, the plant cost is very low.
- Utilities and services – the plant site is near to lots of different water supplies such as a river (Thames) and lakes. As the facility, will have high power and steam requirement, this will be available from various sources such as generators and turbines.

3.2 Plant layout

The detailed design of the layout of the Plant site and Manufacturing facility are given in the appendix. The layout of the manufacturing facilities will be designed in a way to allow a good waste, raw material and personnel flow to minimise the risk and contamination. All of these are an example of current good manufacturing practices which are vital for the production of the therapeutic product (DNA vaccines). The design of manufacturing facility is divided into three sections which include:

- Process Areas – part of facility involve is production process. Includes Fermentation room, alkaline lysis room, purification room, filtration room, raw material room, cell bank, CIP/SIP room and preparation room. (GSK, 2017)
- Process support areas – additional requirements that are important for process operation while following cGMP’s. includes inspection area, a decontamination area, Mobile access lobby, personnel access lobby, ancillary equipment room, packaging and labelling area, conveyor facility. (GSK, 2017)
- Amenities – areas on the site and on the manufacturing facility includes changing rooms, car park, reception desk, security, canteen, etc. (GSK, 2017)

3.21 Layout of the Plant site appendix 1

3.22 Layout of the Production House appendix 2

3.3 Manufacturing Flow

Material/Waste flow

This includes all the movement of materials or waste material. Typical criteria include: (ICheme, 2003)

- Materials flow through the area, for example, linear flow through with no cross-over of production streams.
- Methods of handling and prevention of cross-contamination.
- Storage condition (refrigerated, toxic, hazardous, filtered) (ICheme, 2003)

Raw Materials and supplies flow

Abbildung in dieser Leseprobe nicht enthalten

Personnel Flows

This includes the influence personnel have on the quality of the product that might be caused by their contact with the product. The criteria that have been included in the Production House layout include: (ICheme, 2003)

- Clothing Requirements – (Changing rooms and gowning at the beginning and at each production area).
- Security and access control including potential short cuts and back doors.
- Potential points of cross-contamination between personnel such as changing rooms and gowning room. (ICheme, 2003)

Personnel flow example

Pre-Gowning room to Gowning room (Sterile garments, double gloves) to pDNA production cleanroom.

The layout of the Production house illustrates the Personnel flow, Material flow, waste and equipment flow appendix 2. Different pathways for clean and unclean materials/Personnel to avoid mixing and contamination has been used in the design to comply with cGMP requirements.

3.4 Room Specification data sheets appendix 3

4.0 Plant Services, Systems and Utilities

Plant services, system and utilities are designed to meet the requirement for the manufacturing facility. DNA Vaccine manufacturing requires following services:

Water for CIP to clean equipment and media preparation, Clean steam for SIP to sterilise the equipment, Heating, ventilation and air conditioning (HVAC), water, Compressed air, Effluent treatment, Nitrogen, USP, CIP system, refrigeration and more. (Cole, 1998)

4.1 Heating, ventilation and air-conditioning (HVAC) system

HVAC system is an important part of the production clean rooms design and layout, ensuring that the product purity and quality are not affected by the room temperature, humidity, pressure, air particles and cross contamination. Providing control/healthy environment for process operation and occupants. (UK, 2015)

HVAC system is managed by automated temperature control system (ATC), which controls the humidity, Air distribution and air quality for the process operation. There various types of HVAC systems which are found in the pharmaceutical facilities and selection depends on the requirements within the plant such as environmental conditions and the level of product containment. The degree of control within the plant increases the complexity and the cost of HVAC. (ICheme, 2003)

The HVAC systems for the DNA vaccine manufacturing facility will include: Separate systems for each work centre and total loss systems to minimise the risk of cross-contamination. Terminal HEPA filters on supply and extract to control dust, microorganisms and airborne particles. Sterile (+) or containment (-) pressure cascades by Airlocks and air handling unit (AHU), Low humidity, Dust extract and specified classification of the clean room. (ICheme, 2003)

Abbildung in dieser Leseprobe nicht enthalten

Figure 3: Low humidity total loss containment clean room. Cole, G., Bennett, B. and ICheme, (2003). Engineers guide to pharmaceuticals production - IChemE. Rugby: The Institution of Chemical Engineers.

4.2 Water

The DNA vaccine industry comes under the regulations of MCA, FDA and MHRA. They constantly review with great attention the treatment of water in all segments of the industry used for rinsing, washing and product formulation. (Cole, 1998)

Water is considered to be one of the most difficult product to maintain the required standard not only because of the chemical impurities but, the bacterial contamination, as some system contain components which support the growth of bacteria in water (Cole, 1998). DNA vaccine manufacturing facility will use the water provided by the water treatment plant. Town water, process water and wastewater from the plant will be treated to provide other types of water by various processes such as ion Exchange Deionisation (DI), Reverse Osmosis (RO), Electro Deionisation (EDI) and Distillation (WFI). (Cole, 1998) Types of water will be used in the manufacturing facility are:

Town water/portable water – water from the mains and the quality changes throughout the year. Also, known as the drinking water. Microbiological less than 500cfc/ml and no bacterial colonies according to WHO. (ICheme, 2003)

Process Water – this is normally a portable water which has been passed through the site. Used for washing equipment and cooling. This is also an input for the purified stream (ICheme, 2003)

Purified Water – softened and passed through a UV source to kill live bacteria. Preparation of drugs that do not require WFI (non-sterile products), development of clean solution and input for WFI. According to WHO, Microbiological level less than 100cfc/ml and without any pathogens. (Cole, 1998)

Water for injections (WFI) – softened, extremely low bacterial count and reduced endotoxin. Commonly used for the preparation of the sterile drug. This is generated via distillation with some other treatment method (Cole, 1998). Aerobic Bacteria level < 10cfc/ml and Endotoxin < 0.25 I.U./ml.

The town water and the process water will be used in amenities – admin, lobby, drinking and washing equipment and cooling. The purified water and WFI will be utilised in the main production line for producing the drugs. Purified for initial rinsing and WFI for final rinsing. (GSK, 2017)

Purified water process flow for DNA Vaccine Facility The following process will be used to purify the town/process water for the production of DNA Vaccine.

Abbildung in dieser Leseprobe nicht enthalten

Figure 4: Purified water process flow. Vilhelmsen, B. (2017). Water Systems Challenges for the Pharmaceutical Industry. MDPI Technologist: GSK.

4.3 Clean steam

Clean steam is produced using a steam generator, and it is commonly utilised in a pharmaceutical application where the steam and its condensate are in direct contact with the product to avoid contamination (this is governed by cGMP). (ICheme, 2003)

Clean steam will mainly be used for the sterilisation of process as a sterilising agent; it will also be used in an autoclave and for the humidification for clean rooms. The steam must have the same specification as the water for injections by BP standards for WIF, ensuring the right quality and cleanliness of the steam. This requirement allows the WIF for steam generation. (ICheme, 2003)

Pipeline within the facility can trap condensate which poses the risk of bacterial growth and reduces the effectiveness of sterilisation. Therefore, there should be a steam trap at 30m intervals for efficient removal of condensate. (ICheme, 2003)

4.4 Heat and power

Combined heat & power CHP station will be used onsite to provide heat and power supply. CHP is also known as cogeneration simultaneous production of electrical power and heat from natural gas, and it has a heat recovery system, uses the by-product heat which results in higher thermal efficiency over the conventional power station. The overall efficiency of CHP system is greater than 80%. (admin, 2014)

Therefore, the facility will use CHP station onsite to provide heat and power supply. CHP will operate on natural gas, biomass and waste (as a renewable source). The facility will use the excess thermal energy for steam generation and for heating by using pinch technology.

4.5 Cleaning Systems

It is critical that the DNA vaccine manufacturing facility to obey with all the GMP requirements. The level of hygiene is one of the primary element, involves cleaning plant items and areas that can affect the purity/quality of the manufactured product. Clean in Place (CIP) and sterilise in Place (SIP) will be used in the facility to minimise product loss, removal of soil from product contact surfaces and elimination of micro-organism to meet the desired hygiene standard. But, validation of the cleaning procedure will be necessary before it can be applied. Once this is approved by the relevant governing body, the maintenance of the cleaning systems should be continuously monitored throughout the cleaning and documented which includes the parameters such as flowrate, time, temperature and detergent.

4.51 Clean in Plane (CIP)

CIP is a very simple system, consist of a tank filled with the correct concentration of cleaning agent which is heated to a specified temperature by recirculation and then this is pumped into the pipework or vessel followed by drainage. The important consideration here is the control of flow rate, superficial velocity, temperature and the concentration of detergent of cleaning agent. The facility will use decentralised maximum detergent recovery system. The integration of CIP system with process plant to allow unit cleaning as it has great flexibility, efficiency and objects can be cleaned during production. The cleaning will be measured in different ways such as measurement of mass/volume to assure the right amount of CIP fluid gone through; Pressure ensures that fluid is contacting with the surface at the right force and Conductivity to make sure that the chemical is out of the system. (ICheme, 2003) (Goode, 2017)

Function of typical CIP stages

Abbildung in dieser Leseprobe nicht enthalten

Table 2: Typical function of CIP stages (Goode, 2017)

4.52 Sterilise in Place (SIP)

SIP is not a part of CIP, and this usually comes after the CIP system as detergent cannot clean all part of the plant. The aim of the SIP system is to kill all the viable microorganism (sterilise). Thermal sterilisation (saturated steam sterile) will be used for sterilising the plant equipment which has already been cleaned using CIP system. Similar paraments will be monitored for SIP measurements (Goode, 2017). Steam for cleaning must be pure, and the pressure must be down to 1.2 Barg and temperature 121oC at the point of use.

4.6 Cleanrooms

To produce a sterile product (DNA Vaccine), clean environment is required to minimise the risks of product contamination. In the manufacturing process, people and manufacturing personnel are the biggest sources of contamination, therefore, to reduce the contamination a clean room need to be designed. A clean room is a room in which air supply, air distribution and air filtration are regulated to controls the level of airborne particle concentrates and also meet the cleanliness standards specified by the current standard ISO 14644-1. There is also a specific cleanroom classification for pharmaceutical manufacture figure 4. The table illustrates that the clean rooms are classified into various grades, and it is specified by EU GMP Annex 1. (ICheme, 2003)

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Excerpt out of 37 pages

Details

Title
Plant Design and Manufacturing Principles in DNA Vaccine Production
College
University of Birmingham  (Engineering)
Course
Chemical Engineering
Grade
80.0
Author
Year
2017
Pages
37
Catalog Number
V903166
ISBN (eBook)
9783346248244
ISBN (Book)
9783346248251
Language
English
Tags
Plant Design, Chemical Engineering, DNA Vaccine, Bio Pharmaceutical, engineering, Manufacturing Principles
Quote paper
Sharyar Ahmed (Author), 2017, Plant Design and Manufacturing Principles in DNA Vaccine Production, Munich, GRIN Verlag, https://www.grin.com/document/903166

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