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hello everyone
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in this video we would learn about
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bioprocessing and it's just an overview
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of that
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so what is bioprocessing
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bioprocessing is a technique
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by which we can use biological resources
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such as
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living cells and using their machineries
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we can use products such as enzymes
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metabolites etc which have important
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biomedical relevance now
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definitely the bioprocess would take
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place
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in a bioreactor in a controlled
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environment right
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and the key player of this bioprocessing
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is the biological cell sometimes it
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could be a bacteria sometime it could be
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a mammalian cell
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or sometime it could be a plant cell as
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well so bioprocess
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involves bacteria mammalian cell or even
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cell free systems
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now let's try to understand what type of
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products
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could be generated using this bio
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processing method
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the product could be secreted metabolite
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which is secreted outside
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the product could be an enzyme which is
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relevant for biomedical research or
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maybe this enzyme is
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important therapeutic drug now
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it could be also a recombinant protein
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used for biomedical research so all of
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these type of products can be generated
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using a bio processor now the
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bioprocessor can
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produce these products in a controlled
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environment
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but before we understand bioprocessing
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we need to understand some very basics
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imagine you're cooking for your family
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so you need a very small vessel right
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but
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when you cook for the whole village or
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the whole community you need a bigger
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cooking pot so the environment where
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these
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cooking would happen is different but
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the cooking is same
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the reactions are same but only the
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scale is different
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similarly bioprocessing is also an
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industrial process or it's a macro level
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reaction okay so let's try to take a
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specific example
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so let's say you work with this
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particular enzyme
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and you want this enzyme to be produced
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so what you are going to do
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in lab if you need these enzyme for a
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small scale work
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you would definitely try to generate
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this enzyme
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you would use the cloning workflow where
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you
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clone your gene of interest using the
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cloning methods
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and ultimately you would transfect that
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recombinant plasmid
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into a bacteria this plasmid would
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express
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its product inside the bacteria and you
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would grow the bacteria as the bacteria
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grows the product also grows inside the
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bacteria later on
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using column chromatography and other
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chromatographic tools you would
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purify that protein and finally
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that protein is with you and it
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would be enough for your work but
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imagine
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once you need a some kind of product
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such as an antibody
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such as a restriction enzyme or such as
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recombinant proteins
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for your own research each time
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you are not going to make it from the
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scratch right
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you are going to look for companies who
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deliver
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all of these enzymes right and
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the question is how does these companies
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produce these things in bulk the company
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would deliver these products to your
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doorstep and you would start
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using it but the question is how does
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the bulk preparation takes place
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inside the industry right so we are
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going to talk about that
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industrial aspect of biological
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processing so obviously
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the overall formula is same you need to
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have a recombinant plasmid you need to
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have
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bacteria which would be generating the
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product but
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the reaction container or the reaction
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conditions
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are different and the scale is different
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so you can clearly understand in a
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factory there is a amp up
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in the culture volume from which you are
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purifying
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in lab you might need one microliter of
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this particular enzyme
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but worldwide people need one microliter
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so
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the factory has to generate liters and
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liters of that
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enzyme and that's is not an easy process
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so let's begin from the scratch what is
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happening inside of bioprocessing
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industry
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so they have a seed stock so this seed
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stock would contain
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bacteria you would transfect the
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bacteria with the plasmid
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that you want to express inside this
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bacteria
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now you would gradually scale up your
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culture volume from a small culture
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volume to a big one and ultimately you
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would take your culture
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into a fermentation reactor or a
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bioreactor
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we'll come to the details of the
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bioreactor in a moment
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but in the bioreactor your product would
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be harvested that means your bacteria
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would grow in number as the bacteria is
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growing in number your product is also
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generated
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now after that your product need to be
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recovered from the bacteria right
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so you have to harvest the cells and you
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have to do centrifugation
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followed by some kind of separation by
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chromatographic method
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then there would be several rounds of
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purification steps polishing step
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and ultimately it would pass through a
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quality control
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you have to understand whether the
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product which is generated is actually
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functional or not whether it is okay for
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it to be released in the market so
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quality control step is very important
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and ultimately it would be packaged and
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delivered
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to its proper location so the step
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where the cells are generated in a
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massive amount
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is known as the upstream processing
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so this upstream processes happens in
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the
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bioreactor whereas the recovery process
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purification process
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all of these comprise a downstream
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processing which
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includes processing purification
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polishing quality control and packaging
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so first of all you can take a frozen
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seed stock
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then you can put it in put that culture
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in a
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relatively bigger vessel
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then this culture would be revived
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after that you have to amp up the volume
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so you have to gradually amp up the
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volume
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and once the volume reach quite a lot
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then you have to put it
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in a bioreactor tank so let's talk about
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the bioreactor so there are lots of lots
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of component in this bioreactor tank
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so we are going to talk about it one by
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one so let's look at the bits and pieces
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okay
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so first there is the tank where the
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broth would stay right
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and this tank is actually sterile it
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ensures the microbe that you need
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only is able to grow it does not ensure
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growth of a random microbe obviously
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there is a stirring pedal attached to a
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dc motor so it would allow this pedal to
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rotate
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and its speed can be controlled so in
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this rotation process
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heat might be generated so overall in
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order to maintain the temperature
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of the bioreactor there would be coolant
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systems as well
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so there are coolant systems running
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around these
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broth tank after that there would be a
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aeration unit
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which bubbles oxygen
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through this media now some
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microbes might need oxygen some might
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not need oxygen so this component is
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variable
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so this ensures the dissolved oxygen
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level
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in the bioreactor is in a controllable
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amount
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after that there would be electronic
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display units and there would be control
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boards and control panels there would be
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inlet
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and outlet chamber as well so all that
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comprises bioreactor
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after that the product that is generated
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in the bioreactor would pass through
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centrifuge and this centrifuge does not
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look like the centrifuge
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which is in your lab bench so these
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centrifuge
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are industrial scale centrifuge and they
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are very different looking right
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overall this is a bioreactor and this is
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how the pedals are spinning
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so what we learned so far is the
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upstream process
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the upstream process refers to
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uh the massive amplification of the
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microbes of the cell
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and that would be generating the
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substances that is our
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that is of our interest in a massive
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amount but it still remain
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still remain in an unpurified stage
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right so upstream process can be induced
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upstream process can include um inoculum
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development
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media development improvement of the
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innoculum by genetic engineering process
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so all the molecular biology process at
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the initial step
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and ultimately optimizing the growth
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kinetics
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so you have to come to a growth i mean
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growth condition
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which was which is faster cost effective
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and very efficient so all of these
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things would be
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important when you're making a product
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in an industry skill
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now coming to the downstream process so
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downstream processing refers to
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recovery and purification of the
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biosynthetic product
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which is generated in the upstream
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process now it need to be purified
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so the products that are generated let's
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see how it is purified
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now before that let us take a simple
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example of two kind of scenario
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one type the product could be
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extracellular some kind of metabolite
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which is excreted out of these bacteria
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now the product would be also
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intracellular for example this is the
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enzyme inside the bacteria
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so let's see how each type of products
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can be purified and recovered
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so coming to the extracellular product
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purification
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so in this case the bioreactor tank
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would be connected to a centrifuge and
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this centrifuge the output of the
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centrifuge will be connected to a
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filtration unit so the bio react in the
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bioreactor tank there would be bacteria
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and the bacteria would have several
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metabolites some of these metabolites
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would be of our interest and some are
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useless for us so dur in the centrifuge
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the first pass separation takes place
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so you settle down all the big heavy
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cells in the bottom
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and all the metabolites including useful
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and non useful metabolites are in the
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solution
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now when the solution is passed through
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the filtration unit
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you can purify your product of interest
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based on
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column chromatographic techniques or
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many other techniques so this is how a
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extracellular product can be
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or extracellular metabolite can be
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filtrate
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now coming to a intracellular product
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such as
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an enzyme so let's say this enzyme is
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important for your
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biological research so in order to get
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the enzyme you have to disrupt the cell
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and you need a cell disruption system
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and there are huge
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industrial grade pistons which actually
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breaks apart the cell
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by mechanical shearing and try to get
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all this product out in the solution
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now once the product is out in the
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solution you can so and the product is
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soluble if it's an enzyme
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then you can get a get rid of all of
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these debris using this centrifuge
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whereas my product would be in the
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dissolved state right
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in the supernatant so the supernatant is
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collected and followed by several other
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downstream processing
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such as passing through a column so
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this broth which has our protein of
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interest has to be concentrated because
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there is a huge volume in order of
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thousands of liters of culture could be
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inside a tank
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so that is why this broth need to be
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concentrated in order to recover the
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product
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efficiently now that is why dewatering
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step could be a essential step where you
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remove the excess amount of water
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with the application of vacuum drying
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process
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now there could be initial purification
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of the metabolites for example you use
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hplc based columns or any kind of column
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chromatographic high throughput column
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chromatographic method
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to purify your protein and depending
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upon the nature of your
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product what you want to purify you can
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use your
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columns such as hydrophobic interaction
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column or ion exchange column any kind
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of column you can use in these
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hplc setups again these hplc setups
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differ a lot from
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that we use in our day-to-day lab work
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because everything here is a industrial
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skill process right so it's a macro
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level process lastly there is
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polishing and quality control so this
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polishing polishing ensures that
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this particular product would be 98 to
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100 percent pure and all the purified
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product should be mixed with inert
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ingredients
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and that ensures the product would be
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purified and it would be preserved for a
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long time it won't be degraded quickly
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and ultimately there is a quality
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control step which is very important for
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this product development
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and after these things these products
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would be packaged nicely
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and distributed where where the demand
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is right
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so overall in this video we learned
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about the steps of
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bio bio processing we looked at what is
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downstream what is upstream process we
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kind of had an
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overview of downstream and upstream
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process so in subsequent videos we would
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look at all of these things in nitty
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gritty details but this was just an
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overview
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so i hope you enjoyed this video if you
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