Large Molecules

Introduction

Large molecules are protein-based drugs that can mimic the natural compounds found in the human body. The discovery of these compounds can be done through irrational approach (with actual materials) or rational approach (with virtual simulations).

In Esco Pharma, stirred tank fermenter bioreactor, StirCradle, and StirCradle Pro is capable of culturing and processing bacteria and fungi for research and development purposes to large scale production. Because, of Esco’s biosafety expertise, we offer not only process equipment, but as well as turnkey laboratories and facilities.

Discovery

Pharmaceuticals are new chemical entities (NCEs) and they are produced (synthesized) in manufacturing plants using techniques based on chemical reactions of reactants. Biopharmaceuticals are made using totally different methods. These protein - based drugs are “ manufactured ” in biological systems such as living cells, producing the desired protein molecules in large reaction vessels as the living cells grow, or by extraction from animal serum

The discovery phase for large molecules involves in vitro and in vivo (small animal) studies. When the product proof-of-concept is achieved, process developments to secure Investigational New Drug (IND) is planned.

CelCradle™ is a disposable bioreactor capable of high-density cell culture for protein expression, virus, and monoclonal antibody production. It is designed based on the concept of bellow-induced intermittent flow of media and air through porous matrices, where cells reside. This provides a low shear, high aeration, and foam-free culture environment.

 

Most companies partner with Contract Development Manufacturing Organizations (CDMOs) to help develop and manufacture the drug as well as offer Quality and Regulatory support, mostly for large molecules, hence the increasing manufacturing cost and stricter requirements.

Aster BioPharma works with partner companies that are utilizing VacciXcell’s Tide Motion Platform to provide CDMO services for them in the Far East allowing them to gain market access into the region. Aster BioPharma also works with top academic institutes to translate research into clinical products and provide custom services to start ups and large-scale biotechnology companies.

CDMO services running on Esco’s VacciXcell are now provided by a new independent company called Esco Aster Biopharma operating in #07-26, 71 Ayer Rajah Crescent, Singapore 139951

 

Clinical Trial

Upstream manufacturing  includes operations related to cell expansion steps starting with a single vial of frozen cells and growing these exponentially into larger and larger systems eventually reaching your large-scale terminal reactor where the targeted protein is expressed.

Downstream manufacturing, or commonly referred to as purification, is focused on the capture and isolation of a targeted molecule and the removal of impurities. This is accomplished through several different processes to include filtration, column chromatography, and tangential flow filtration (TFF).

 

Commercial Production/Delivery 

The facility and the utility requirements are the fundamental support of the process flow and production effectiveness. In facility design, it is important to consider the regulatory nature of the industry and seeking some in-depth knowledge of the requirements, hence, serving the most basic foundation for a safe, effective, and pure drugs while maintain the cost.

Large volumes of air are required to properly satisfy the international standards for clean room air. Air handling systems such as isolators and the built and design of the cleanroom is critical as these will edict the permitted processes during the sterile compounding.

Cleanrooms are extremely critical on the manufacturing of biologics, hence the used of Cross Contamination Facility Integrated Barrier (CCFIB) can maximize the performance given by a cleanroom to ensure the purity of any drug product.   This equipment will minimize the exposure of chemical or biological contaminants, such as micro-organisms or adventitious agents, thus providing utmost protection to the product, operator and the environment.

The CradlePro-Iso is an integrated system that combines the Esco Versati™ Centrifuge, Esco CO2 incubator, and CelCradle™ benchtop bioreactor system inside the Esco HPI G3. The CelCradle™ system provides cells with an environment of low shear stress, zero foaming, and of high rates of aeration and nutrition, while the CO2 incubator controls the cells surrounding conditions.

The CradlePro-Iso was designed to fully enclose the cell processing system inside an isolator to ensure the safety of the operator, without compromising the quality of the product.

 

RABS and isolators can be used in the manufacture of biologics, including vaccines, gene therapies, and protein-based drugs. Often, biologic products are preservative-free, contain growth media, and are easily susceptible to contamination. Another area that demands the use of RABS and isolators is the manufacture of sterile drug products with toxic, cytotoxic, and highly potent molecules, which require stringent barriers to protect personnel who are handling these materials. In general, RABS and isolators are being used for smaller-volume and high-value pharmaceuticals. The benefit/cost balance has to be considered when discussing the use of barriers: RABS and isolators come with a high price tag and are associated with additional expenses related to the operation of a cleanroom, such as energy costs, operating costs, testing costs, and gown costs.

Additional cGMP requirements are also included for the commercial production and delivery of large molecules. These requirements are but not limited to: cleanliness, validation and commissioning, process-material personnel and air flows, cold storage, dry storage, emergency procedures, safety, security, pest control, environmental monitoring, preventive maintenance and calibrations, and all possible redundancies that may be required to support utility and processes.

Fill/Finish Operations

The fill/finish team is a specialized function within the manufacturing team dedicated to the manufacture of the drug product. A drug product refers to the final formulated product in its delivery container. In most cases for biologic products, these will be traditional glass vials. Other systems include prefilled syringes or intravenous (IV) bags. As these steps in the process are the last manufacturing.

Capping Machine

Depyrogenation Tunnel

External Vial Washer

Filling/Stopper Inserting Machine Trayloader Vial Washer

There are two distinct technologies dominating the fill/finish process: isolators and RABS. Each technology has its advantages. With isolator technology, the processing takes place in systems that are entirely shut off from the outside environment. As it pertains to sterility assurance levels (SAL), isolators are often considered the best solution due to the automatic decontamination processes involved. However, isolators need extensive decontamination and preparation processes following a batch to enable a safe change in product.

RABS technology also achieves the SAL currently required by regulatory authorities. With this technology, the physical barriers of a production plant are limited; a RABS requires installation in a higher-class environment (at least ISO 7, with the RABS located in an ISO 5 area). (Hernandez, 2017, Best Practices for Sterility Assurance in Fill/Finish Operations)

Quality Assurance

Quality assurance provides a fully independent look at all documentation, production areas, and supply chain functions to ensure compliance. Quality is responsible for ensuring all raw materials, procedures, and areas are released for production. They will also perform reviews of all documentation, quality control samples, and final release specifications before approving a batch for release.

In-Process Tests

For unprocessed bulk harvest common in-process tests are:

• Bioburden

• Mycoplasma detection with mycoplasmastasis

• In vitro assay for nonendogenous or adventitious viruses

• Detection of Mouse Minute Virus (MMV) DNA by quantitative polymerase chain reaction (qPCR),

• Transmission electron microscopy (TEM) of supernatant Endotoxin testing is performed on the clarified bulk harvest

Release Tests for Drug Substance

• Bioburden

• Endotoxin

Quality Control

The quality control unit is responsible for performing testing on raw materials, in-process and final product testing, and environmental monitoring activities in a biologics facility.

The compendial sterility test is the most common nominal test for the quality control of large molecules which describes two separate types of approach. In the first method, a solution from a specified number of containers (volume and number determined by batch size and unit fill volume) is filtered through a filter of nominal pore size 0.45 μm. Recovery of viable cells from the filter(s) is performed by submerging the filter in one of two recovery media followed by incubation at specified temperatures for 14 days. The second test is a direct immersion of the product or suspensions into a suitable volume of the two media to allow growth. The media are designed to support growth in aerobic, or growth in an environment of limited oxygen availability. This test requires demonstration that the specific method used is suitable for that product. In this case, a sterility test isolator is needed.

Facilities and Engineering

The facilities and engineering teams oversee all of the key systems required to keep the manufacturing plant operational. This includes base building systems, process equipment, utilities, building automation, and heating, ventilation, and air conditioning (HVAC). They also perform routine and nonroutine maintenance activities on the aforementioned systems. These teams are staffed with skilled.

CDMO Services vs. In-House Manufacturing

Most CDMOs have an already built-in facility that can cater all the processes needed by large molecules from the research and development to the QC and IPQC processes. These CDMOs have the capability to to produce and purify biologic drugs at a certain scale to accommodate their scale or “niche” market. These capabilities are large capital expenditures such as stainless steel bioreactors, fermentors, mixing tanks, water systems, chromatography skids, chromatography columns, cleanrooms, and QC laboratories. The advantage of a CMO facility is that it is ready for manufacturing the developed product.

A major disadvantage for the in-house model is the capital outlay for an in-house facility. In terms of time and equipment, an in-house facility would be very costly in respect to supporting in-house expertise to manage the design, fabrication, installation and qualification of the facility, process, and support equipment. There are other considerations such as cleaning and maintenance of all product contact surfaces such as bioreactors and mixing tanks. These require studies and validation of the cleaning processes. Samples would have to be taken and tested creating additional in-house expertise, time, and resources and this would be contingent on suite availability to develop and manufacture the product. Whereas outsourcing these activities to a contract development manufacturing organization (CDMO) would shorten the timeline significantly because the CMO would already have the expertise, process, and facility equipment in place to support the development and manufacturing activities.

Aster BioPharma works with partner companies that are utilizing VacciXcell’s Tide Motion Platform to provide CDMO services for them in the Far East allowing them to gain market access into the region. Aster BioPharma also works with top academic institutes to translate research into clinical products and provide custom services to start ups and large-scale biotechnology companies.

CDMO services running on Esco’s VacciXcell are now provided by a new independent company called Esco Aster Biopharma operating in #07-26, 71 Ayer Rajah Crescent, Singapore 139951