Bio-processing overview (Upstream and downstream process)
Summary
TLDRThis video offers an insightful overview of bioprocessing, a technique that harnesses biological resources like living cells to produce biomedically relevant products such as enzymes and recombinant proteins. It explains the process taking place in bioreactors, involving various cell types, and differentiates between upstream and downstream processing. Upstream focuses on cell growth and product generation, while downstream encompasses recovery, purification, and quality control of the synthesized products. The video also touches on industrial-scale challenges and the equipment used, such as centrifuges and chromatography systems, highlighting the transition from lab-scale to bulk production in the bioprocessing industry.
Takeaways
- 🧪 Bioprocessing is a technique that utilizes biological resources such as living cells to produce enzymes, metabolites, and other products with biomedical relevance.
- 🌱 The process takes place in a bioreactor, which provides a controlled environment for the biological cells to function, and can involve bacteria, mammalian cells, or plant cells.
- 🛠️ Bioprocessing is an industrial-scale process, differentiated from lab-scale by the scale of the reaction, not the type of reaction.
- 🔬 Products generated through bioprocessing can include secreted metabolites, enzymes for research or therapeutic drugs, and recombinant proteins for biomedical research.
- 🏭 The industrial production of biological products involves scaling up from a small culture volume to a large bioreactor, where the product is generated in bulk.
- 🌱 The bioprocessing industry starts with a seed stock of bacteria, which is then transfected with a plasmid and scaled up to produce the desired product.
- 🔄 The process involves both upstream and downstream processing; upstream refers to cell growth and product generation, while downstream involves recovery, purification, and packaging of the product.
- 🌡️ Bioreactors are equipped with various components including a sterile tank, stirring mechanism, temperature control, and aeration to ensure optimal conditions for cell growth.
- 🔬 Centrifuges and filtration units are used in the downstream process to separate cells from the product and to purify the product based on its location (intracellular or extracellular).
- 🔄 Cell disruption systems are used for intracellular products to release the product from within the cells, followed by purification steps such as chromatography.
- 🏷️ Quality control is a critical step in bioprocessing to ensure the functionality and safety of the product before it is released for use.
Q & A
What is bioprocessing?
-Bioprocessing is a technique that utilizes biological resources, such as living cells, to produce products like enzymes and metabolites with biomedical relevance in a controlled bioreactor environment.
What are the types of biological cells that can be used in bioprocessing?
-The biological cells used in bioprocessing can be bacteria, mammalian cells, or plant cells.
What are some examples of products generated using bioprocessing?
-Examples of products generated using bioprocessing include secreted metabolites, enzymes for biomedical research, therapeutic drugs, and recombinant proteins.
What is the purpose of a bioreactor in bioprocessing?
-A bioreactor is used to carry out the bioprocess in a controlled environment, allowing for the growth of biological cells and the production of the desired products.
How does the scale of bioprocessing differ from a small-scale lab experiment to an industrial process?
-In a lab, a small vessel is used for a small scale work, whereas in an industrial process, a much larger bioreactor is used to produce the product in bulk, catering to a larger demand.
What is the role of a seed stock in bioprocessing?
-A seed stock contains bacteria that are transfected with the desired plasmid. It serves as the starting point for gradually scaling up the culture volume in bioprocessing.
What are the components of a bioreactor?
-A bioreactor consists of a sterile tank for the broth, a stirring paddle attached to a motor, a coolant system to maintain temperature, an aeration unit for oxygen supply, and electronic control systems.
What is the difference between upstream and downstream processing in bioprocessing?
-Upstream processing involves the generation of cells in large amounts and the production of the product, while downstream processing includes the recovery, purification, polishing, quality control, and packaging of the product.
How is an extracellular product purified and recovered in bioprocessing?
-An extracellular product is purified and recovered by connecting the bioreactor tank to a centrifuge and then to a filtration unit, which separates the cells from the metabolites and purifies the product of interest using column chromatographic techniques.
How is an intracellular product such as an enzyme recovered in bioprocessing?
-An intracellular product is recovered by first disrupting the cells using a cell disruption system, which mechanically shears the cells to release the product. The product is then separated from the debris using centrifugation and further purified through downstream processing steps.
What is the importance of quality control in bioprocessing?
-Quality control is crucial in bioprocessing to ensure that the final product is functional, meets the required specifications, and is safe for release into the market.
Outlines
🌱 Introduction to Bioprocessing
This paragraph introduces the concept of bioprocessing, a technique that utilizes biological resources such as living cells to produce biomedically relevant products like enzymes and metabolites. It explains that bioprocessing occurs in a controlled environment known as a bioreactor and involves biological cells, which can be bacteria, mammalian, or plant cells. The paragraph also distinguishes between different types of products generated through bioprocessing, such as secreted metabolites, enzymes, and recombinant proteins. The speaker uses an analogy of cooking to explain the scale difference between laboratory and industrial bioprocessing and sets the stage for a deeper dive into the industrial aspect of bioprocessing, including the production of enzymes in bulk by companies.
🔬 Bioprocessing Industry Overview
The second paragraph delves into the specifics of the bioprocessing industry, starting with the seed stock containing bacteria that are transfected with a plasmid to express the desired product. It outlines the process of scaling up the culture volume and the use of a bioreactor for product harvesting. The paragraph describes the downstream processing, which includes cell harvesting, centrifugation, separation, purification, and quality control, emphasizing the importance of each step in ensuring the product's functionality and market readiness. The speaker also explains the components of a bioreactor, including the tank, stirring mechanism, aeration unit, and control systems, and discusses the industrial-scale centrifuges used in the process. This paragraph provides a comprehensive overview of the bioprocessing steps, from upstream processing in the bioreactor to downstream processing for product recovery and purification.
🛠️ Purification and Recovery of Bioproducts
The third paragraph focuses on the purification and recovery of bioproducts, distinguishing between extracellular and intracellular products. For extracellular products, the process involves centrifugation to separate cells from metabolites, followed by filtration and chromatographic techniques to purify the product of interest. In contrast, intracellular products require cell disruption systems to release the product, such as enzymes, into solution. The paragraph explains the use of industrial-grade pistons for mechanical shearing and the subsequent steps of purification, including dewatering, concentration, and chromatographic methods like HPLC. The importance of polishing steps to achieve high purity and quality control to ensure product functionality and preservation is highlighted. The paragraph concludes with the packaging and distribution of the final bioproducts, providing a complete picture of the bioprocessing journey from raw materials to finished goods.
Mindmap
Keywords
💡Bioprocessing
💡Bioreactor
💡Biological Cell
💡Metabolites
💡Enzymes
💡Recombinant Proteins
💡Upstream Processing
💡Downstream Processing
💡Cell Disruption
💡Quality Control
💡Industrial Scale
Highlights
Bioprocessing is a technique that uses biological resources like living cells and their machinery to produce biomedically relevant products.
Bioreactors are used in bioprocessing to provide a controlled environment for biological cells to produce products.
The key player in bioprocessing is the biological cell, which can be bacteria, mammalian, or plant cells.
Bioprocessing can generate secreted metabolites, enzymes, and recombinant proteins for biomedical research and therapeutic drugs.
An analogy is made between cooking for a family versus a community to explain the scale differences in bioprocessing.
In labs, enzymes are produced through cloning and transfection into bacteria, which is then scaled up for industrial production.
Companies produce enzymes and proteins in bulk using bioprocessing techniques for research and therapeutic use.
The bioprocessing industry starts with a seed stock of bacteria that is transfected with a plasmid to express the desired product.
Culture volume is gradually scaled up from small to large in bioreactors for the production of large quantities of the product.
Products are harvested from the bacteria through centrifugation and chromatographic separation methods.
Quality control is crucial to ensure the functionality and safety of the bioprocessed products before market release.
Upstream processing involves the generation of cells in large amounts in bioreactors, while downstream processing includes recovery, purification, and packaging.
Bioreactors consist of a sterile tank, stirring paddle, DC motor, coolant system, aeration unit, and electronic control units.
Industrial-scale centrifuges are used to separate cells and metabolites in the bioprocessing of extracellular products.
Intracellular products require cell disruption systems to release the product, followed by purification steps.
Concentration and dewatering steps are essential for efficiently recovering products from large volumes of culture broth.
High-throughput column chromatographic methods such as HPLC are used for the initial purification of the product.
Polishing steps ensure the product is 98 to 100 percent pure, and quality control is vital for product development.
The video provides an overview of bioprocessing, with subsequent videos diving into more detailed aspects of the process.
Transcripts
hello everyone
in this video we would learn about
bioprocessing and it's just an overview
of that
so what is bioprocessing
bioprocessing is a technique
by which we can use biological resources
such as
living cells and using their machineries
we can use products such as enzymes
metabolites etc which have important
biomedical relevance now
definitely the bioprocess would take
place
in a bioreactor in a controlled
environment right
and the key player of this bioprocessing
is the biological cell sometimes it
could be a bacteria sometime it could be
a mammalian cell
or sometime it could be a plant cell as
well so bioprocess
involves bacteria mammalian cell or even
cell free systems
now let's try to understand what type of
products
could be generated using this bio
processing method
the product could be secreted metabolite
which is secreted outside
the product could be an enzyme which is
relevant for biomedical research or
maybe this enzyme is
important therapeutic drug now
it could be also a recombinant protein
used for biomedical research so all of
these type of products can be generated
using a bio processor now the
bioprocessor can
produce these products in a controlled
environment
but before we understand bioprocessing
we need to understand some very basics
imagine you're cooking for your family
so you need a very small vessel right
but
when you cook for the whole village or
the whole community you need a bigger
cooking pot so the environment where
these
cooking would happen is different but
the cooking is same
the reactions are same but only the
scale is different
similarly bioprocessing is also an
industrial process or it's a macro level
reaction okay so let's try to take a
specific example
so let's say you work with this
particular enzyme
and you want this enzyme to be produced
so what you are going to do
in lab if you need these enzyme for a
small scale work
you would definitely try to generate
this enzyme
you would use the cloning workflow where
you
clone your gene of interest using the
cloning methods
and ultimately you would transfect that
recombinant plasmid
into a bacteria this plasmid would
express
its product inside the bacteria and you
would grow the bacteria as the bacteria
grows the product also grows inside the
bacteria later on
using column chromatography and other
chromatographic tools you would
purify that protein and finally
that protein is with you and it
would be enough for your work but
imagine
once you need a some kind of product
such as an antibody
such as a restriction enzyme or such as
recombinant proteins
for your own research each time
you are not going to make it from the
scratch right
you are going to look for companies who
deliver
all of these enzymes right and
the question is how does these companies
produce these things in bulk the company
would deliver these products to your
doorstep and you would start
using it but the question is how does
the bulk preparation takes place
inside the industry right so we are
going to talk about that
industrial aspect of biological
processing so obviously
the overall formula is same you need to
have a recombinant plasmid you need to
have
bacteria which would be generating the
product but
the reaction container or the reaction
conditions
are different and the scale is different
so you can clearly understand in a
factory there is a amp up
in the culture volume from which you are
purifying
in lab you might need one microliter of
this particular enzyme
but worldwide people need one microliter
so
the factory has to generate liters and
liters of that
enzyme and that's is not an easy process
so let's begin from the scratch what is
happening inside of bioprocessing
industry
so they have a seed stock so this seed
stock would contain
bacteria you would transfect the
bacteria with the plasmid
that you want to express inside this
bacteria
now you would gradually scale up your
culture volume from a small culture
volume to a big one and ultimately you
would take your culture
into a fermentation reactor or a
bioreactor
we'll come to the details of the
bioreactor in a moment
but in the bioreactor your product would
be harvested that means your bacteria
would grow in number as the bacteria is
growing in number your product is also
generated
now after that your product need to be
recovered from the bacteria right
so you have to harvest the cells and you
have to do centrifugation
followed by some kind of separation by
chromatographic method
then there would be several rounds of
purification steps polishing step
and ultimately it would pass through a
quality control
you have to understand whether the
product which is generated is actually
functional or not whether it is okay for
it to be released in the market so
quality control step is very important
and ultimately it would be packaged and
delivered
to its proper location so the step
where the cells are generated in a
massive amount
is known as the upstream processing
so this upstream processes happens in
the
bioreactor whereas the recovery process
purification process
all of these comprise a downstream
processing which
includes processing purification
polishing quality control and packaging
so first of all you can take a frozen
seed stock
then you can put it in put that culture
in a
relatively bigger vessel
then this culture would be revived
after that you have to amp up the volume
so you have to gradually amp up the
volume
and once the volume reach quite a lot
then you have to put it
in a bioreactor tank so let's talk about
the bioreactor so there are lots of lots
of component in this bioreactor tank
so we are going to talk about it one by
one so let's look at the bits and pieces
okay
so first there is the tank where the
broth would stay right
and this tank is actually sterile it
ensures the microbe that you need
only is able to grow it does not ensure
growth of a random microbe obviously
there is a stirring pedal attached to a
dc motor so it would allow this pedal to
rotate
and its speed can be controlled so in
this rotation process
heat might be generated so overall in
order to maintain the temperature
of the bioreactor there would be coolant
systems as well
so there are coolant systems running
around these
broth tank after that there would be a
aeration unit
which bubbles oxygen
through this media now some
microbes might need oxygen some might
not need oxygen so this component is
variable
so this ensures the dissolved oxygen
level
in the bioreactor is in a controllable
amount
after that there would be electronic
display units and there would be control
boards and control panels there would be
inlet
and outlet chamber as well so all that
comprises bioreactor
after that the product that is generated
in the bioreactor would pass through
centrifuge and this centrifuge does not
look like the centrifuge
which is in your lab bench so these
centrifuge
are industrial scale centrifuge and they
are very different looking right
overall this is a bioreactor and this is
how the pedals are spinning
so what we learned so far is the
upstream process
the upstream process refers to
uh the massive amplification of the
microbes of the cell
and that would be generating the
substances that is our
that is of our interest in a massive
amount but it still remain
still remain in an unpurified stage
right so upstream process can be induced
upstream process can include um inoculum
development
media development improvement of the
innoculum by genetic engineering process
so all the molecular biology process at
the initial step
and ultimately optimizing the growth
kinetics
so you have to come to a growth i mean
growth condition
which was which is faster cost effective
and very efficient so all of these
things would be
important when you're making a product
in an industry skill
now coming to the downstream process so
downstream processing refers to
recovery and purification of the
biosynthetic product
which is generated in the upstream
process now it need to be purified
so the products that are generated let's
see how it is purified
now before that let us take a simple
example of two kind of scenario
one type the product could be
extracellular some kind of metabolite
which is excreted out of these bacteria
now the product would be also
intracellular for example this is the
enzyme inside the bacteria
so let's see how each type of products
can be purified and recovered
so coming to the extracellular product
purification
so in this case the bioreactor tank
would be connected to a centrifuge and
this centrifuge the output of the
centrifuge will be connected to a
filtration unit so the bio react in the
bioreactor tank there would be bacteria
and the bacteria would have several
metabolites some of these metabolites
would be of our interest and some are
useless for us so dur in the centrifuge
the first pass separation takes place
so you settle down all the big heavy
cells in the bottom
and all the metabolites including useful
and non useful metabolites are in the
solution
now when the solution is passed through
the filtration unit
you can purify your product of interest
based on
column chromatographic techniques or
many other techniques so this is how a
extracellular product can be
or extracellular metabolite can be
filtrate
now coming to a intracellular product
such as
an enzyme so let's say this enzyme is
important for your
biological research so in order to get
the enzyme you have to disrupt the cell
and you need a cell disruption system
and there are huge
industrial grade pistons which actually
breaks apart the cell
by mechanical shearing and try to get
all this product out in the solution
now once the product is out in the
solution you can so and the product is
soluble if it's an enzyme
then you can get a get rid of all of
these debris using this centrifuge
whereas my product would be in the
dissolved state right
in the supernatant so the supernatant is
collected and followed by several other
downstream processing
such as passing through a column so
this broth which has our protein of
interest has to be concentrated because
there is a huge volume in order of
thousands of liters of culture could be
inside a tank
so that is why this broth need to be
concentrated in order to recover the
product
efficiently now that is why dewatering
step could be a essential step where you
remove the excess amount of water
with the application of vacuum drying
process
now there could be initial purification
of the metabolites for example you use
hplc based columns or any kind of column
chromatographic high throughput column
chromatographic method
to purify your protein and depending
upon the nature of your
product what you want to purify you can
use your
columns such as hydrophobic interaction
column or ion exchange column any kind
of column you can use in these
hplc setups again these hplc setups
differ a lot from
that we use in our day-to-day lab work
because everything here is a industrial
skill process right so it's a macro
level process lastly there is
polishing and quality control so this
polishing polishing ensures that
this particular product would be 98 to
100 percent pure and all the purified
product should be mixed with inert
ingredients
and that ensures the product would be
purified and it would be preserved for a
long time it won't be degraded quickly
and ultimately there is a quality
control step which is very important for
this product development
and after these things these products
would be packaged nicely
and distributed where where the demand
is right
so overall in this video we learned
about the steps of
bio bio processing we looked at what is
downstream what is upstream process we
kind of had an
overview of downstream and upstream
process so in subsequent videos we would
look at all of these things in nitty
gritty details but this was just an
overview
so i hope you enjoyed this video if you
like this video don't forget to sh
subscribe to my channel hit that bell
icon for notification share this video
in the social media for
such that i can reach bigger audience my
lectures are also present in an academy
which is india's biggest online learning
platform
and you can use ap 10 my code to get a
10
discount in my courses thank you guys
Weitere ähnliche Videos ansehen
5.0 / 5 (0 votes)