Search results
Learn about the stages and methods of upstream and downstream processing in industrial fermentation, such as fermentation medium, inoculum, fermentation systems, and product recovery. This lesson is part of a course on biotechnology and bioprocessing by IASRI.
- Objectives
- Overview
- Dispensing room
- CIP/SIP systems
- Media preparation area
- Conductivity
- Cell Culture/Fermentation
- Environment necessary for cell growth
- Figure 10-3. Aseptic preparation of cell bank vials in a Biological Safety Cabinet (BSC)
- Inoculum
- Bioreactors
- Stainless steel bioreactors
- Production strategy
- Bioreactor preparation
- Figure 10-18. Tube welding
- Figure 10-19. Tube sealing
- PV = nRT
- Cell counting
- Primary recovery (harvest)
- Figure 10-24. A centrifuge, motor and associated piping
- Figure 10-25 A depth filter housing and internal filter stack
- Filter integrity testing
- Check Your Knowledge
- GeneratedCaptionsTabForHeroSec
This chapter provides an overview of the key elements for operation of a biomanufacturing upstream processing production area. Upon completion of this chapter the student will be able to: distinguish between mammalian and microbial upstream processing describe the contribution of the areas of dispensing, media preparation, and fermentation/cell cul...
The upstream biomanufacturing technician’s tasks center around microscopic, living cells. The main objective of upstream manufacturing is to create the environment necessary for cells to make the target protein. These proteins serve various medicinal purposes. Biomanufacturing is used to produce products such as therapeutic proteins, antibiotics, h...
Materials needed by the biomanufacturing process to make product are weighed and measured in a traceable, controlled, and sanitary area prior to their use. Optimally, raw materials are received into the manufacturing facility in packaging which is suitable for cGMP processing areas (e.g., glass or plastic with no cardboard or other fiber shedding m...
Vessels used during the biomanufacturing process and all associated piping/hoses must be free of any foreign substances prior to use. Foreign substances include cell debris, medium, cleaning chemicals, and even the target protein from a prior batch. As most bioreactors are multi-use (and may be multi-product), any substances inadvertently left behi...
Technicians working in media preparation prepare media with precise recipes required for each bioreactor stage of scale-up from inoculum (thawed cells introduced to the media) to harvest, where cells are separated from the media and the target API. As a human body requires a certain amount of carbohydrates, fats (lipids), and proteins in a diet to ...
Conductivity is a measure of the ions dissociated in the solutions. Conductivity measurements are important in CIP, as they reveal the purity of the rinse water. CIP cleaning uses acids and bases to clean tanks and is followed by rinsing with WFI. Conductivity measures the level of any residual acid or base ions left in the rinse water-the higher t...
The goal of upstream biomanufacturing is to grow cells that will produce a desired protein, the protein being further purified during the downstream processing steps (see Downstream Processing chapter). The stages within the upstream biomanufacturing area are generally referred to as inoculum, bioreactor stage (seed and production), and primary rec...
All living things grow by cellular division, and the same applies to fermentation or cell culture - the cells divide and continue to multiply. Just as humans need certain things to continue to live (food, water, O2, a specific body temperature, etc.), cells in culture settings also require a specific environment to support growth. The upstream biom...
A cell bank should have a full history of the cell line, as well as the criteria used to determine its authenticity (e.g., test results showing absence of bacterial, fungal, viral, and Mycoplasma contamination). Test results should also demonstrate cell line characterization, including isoenzyme analysis, chromosome analysis, and in some cases, DNA...
The inoculum stage involves the thawing of a frozen vial of cells (ampoule) and its introduction into a bioreactor. Placing cells in the bioreactor allows for scaling-up of the culture to meet cell density and volume requirements in order to inoculate the next stage in the process. The inoculum stage is comparable to the childhood stage in the prod...
The next stage in the production process for protein-based therapeutics is the use of seed and production bioreactors to create the conditions that optimize growth of the mammalian cells. Bioreactors are classified as either disposable or stainless steel. Regardless, all use the same basic principles in operation and use.
This type of a bioreactor vessel is a double-walled, glycol-jacketed, sanitary pressure vessel designated to provide primary containment for the cell culture. There are four layers to the stainless steel bioreactor vessel: Interior wall provides the sterile product contact surface for the cell culture made of stainless steel (316L) and designed acc...
The manufacturing process step involving bioreactors is further classified by the type of production strategy: Batch: the culture generally goes through the growth cycle of lag, exponential, plateau, and death; the production cycle is short in this type of strategy; protein is harvested one time at the end of the production cycle. Fed batch: the cu...
In the case of disposable bioreactors, a cell bag is sterilized (using Gamma irradiation) and set up to receive media and cells for cell culture. For stainless steel bioreactors, the bioreactor is prepared by cleaning and sterilizing using CIP and SIP. Leak detection is usually performed by pressure-testing the vessel with helium. The biomanufactur...
When either additions are being made or samples are being removed from wave bags, bioreactors, harvest tanks, VI tanks, hold tanks, etc., tube welding is the general method used for connecting the addition/ sample container to the vessel. Tube welders are validated per cGMP industry standards. A Sterile Tube Fuser uses a non-reusable blade to cut, ...
After additions are performed or samples are pulled via tubing, tube sealing is the general method used for separating the associated containers/vessels. The tube sealer, validated per cGMP industry standards, is used to permanently seal tubing. This is done by clamping the tubing in a precise position between two heated jaws. The molten tubing is ...
(where P = Pressure, V = Volume, T = Temperature in degrees Kelvin) n = moles of gas, R = Gas Constant, This relationship, along with the ability of the meter to compensate for temperature changes, allows results to be expressed as percentage oxygen or mg/L O2.
As the cells grow during the biomanufacturing process, it is necessary to keep track of cell growth. Cell counting tallies the number of viable (living) and non-viable (dead) cells and calculates a viability percentage. This percentage is the number of living cells as a percentage to the total number of cells (living and dead) in the culture. Viabl...
The main objective of primary recovery, or harvest, is to separate the cells from the media containing the target API. It is the first step in recovery of the protein product from the culture. Each cell line has been studied, and depending on the batch record, there are certain triggers on the proper time to end the cell culture/fermentation proces...
The next step in cell harvest is the filtration step to remove large debris. The most obvious function of a filter is that of mechanical straining, or size exclusion. Particles too large to pass through the filter membrane are either trapped on the surface or within pore channels of what is called the tortuous path. In sterile biomanufacturing, ven...
Depth filtration can also exploit charge difference between what is removed and the filter matrix itself. Electrokinetic adsorption results in positively-charged filter matrix components, attaching to negatively-charged items (bacteria, Mycoplasma, viruses, yeast, and endotoxins). These adhere to the filter and are firmly retained. The resulting po...
To ensure filtration meets the intended purpose, filter integrity testing is performed during the biomanufacturing operation. The purpose of filter integrity testing is to ensure the integrity of the filter element(s) and the proper installation of the filter element(s). There are several types of filter integrity tests. Pre-use integrity testing d...
Chemicals that are used in CIP are usually dissolved to produce a strong base solution. An example is . The exponential growth phase of cells is called: lag phase log phase peak phase plateau phase Cells can be grown in inoculum using a variety of equipment. Depending on the type of cell line, the culture may be or in suspension. If pH is low it ...
Learn the key elements and objectives of upstream processing in biomanufacturing, such as dispensing, media preparation, fermentation, and cell culture. Find definitions and examples of terms related to upstream processing, such as bioreactor, centrifugation, and CIP.
Learn the definitions and examples of downstream and upstream processing in biotechnology. Downstream processing is the recovery and purification of products from natural sources, while upstream processing is the cultivation and harvesting of cells or microbes.
Jan 1, 2018 · Developing a robust, high-yielding upstream process is a complex and challenging task, and a good result is a combination of powerful technology, high-quality starting materials, and clever process design.
- Eva K. Lindskog
- 2018
Nov 24, 2021 · Learn the difference between upstream and downstream processing in biopharmaceutical production. Upstream processing involves cell cultivation and harvest, while downstream processing involves product purification and concentration.
Sep 1, 2019 · Textbook definition: Upstream processing refers to the stage of bioprocessing where cells are grown to the desired quantity in bioreactors, and all stages related to this such as cell isolation, cell cultivation, media preparation, cell banking & storage to culture expansion until harvest.
New to process development for biomanufacturing? Here are definitions, activities, and things to consider when developing an upstream or downstream process.
