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Question
Which of the following methods does not help in removal of cellular debris from the culture media during downstream processing of the recombinant protein?
Which of the following methods does not help in removal of cellular debris from the culture media during downstream processing of the recombinant protein?
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Solution
The correct option is C Electroporation
Genetically modified cells are cultured at a large scale in specially designed bioreactors to have large scale yield of the desired recombinant proteins. Once the synthesis of the proteins is over, they are separated from the medium and purified before being marketed for commercial use. This is known as downstream processing.
The very first step of downstream processing involves separation of the insoluble cellular debris and other particulate matter from the culture medium containing the recombinant protein. This can be achieved using several techniques such as filtration, centrifugation, sedimentation, etc.
In filtration technique, a filtration matrix or membrane is used to separate the culture medium from the cellular biomass. For example, bacterial cells can be filtered out using membrane filters that have pore size smaller than that of the bacteria.
Centrifugation helps in separation of components in a mixture based on the difference in the densities. Due to the high centrifugal force, the denser cellular debris settles down at the bottom, leaving the culture medium containing the soluble protein at the top.
Sedimentation is a method in which the mixture of culture medium is kept undisturbed for a long period. The heavier cellular debris settles at the bottom. The culture medium with the recombinant protein becomes the supernatant.
Electroporation is a technique used for introduction of foreign DNA into bacterial or plant cells by subjecting them to a very high voltage or electrical pulse. This results in creation of transient pores in the cell membrane through which the foreign DNA can easily enter the cells.
Genetically modified cells are cultured at a large scale in specially designed bioreactors to have large scale yield of the desired recombinant proteins. Once the synthesis of the proteins is over, they are separated from the medium and purified before being marketed for commercial use. This is known as downstream processing.
The very first step of downstream processing involves separation of the insoluble cellular debris and other particulate matter from the culture medium containing the recombinant protein. This can be achieved using several techniques such as filtration, centrifugation, sedimentation, etc.
In filtration technique, a filtration matrix or membrane is used to separate the culture medium from the cellular biomass. For example, bacterial cells can be filtered out using membrane filters that have pore size smaller than that of the bacteria.
Centrifugation helps in separation of components in a mixture based on the difference in the densities. Due to the high centrifugal force, the denser cellular debris settles down at the bottom, leaving the culture medium containing the soluble protein at the top.
Sedimentation is a method in which the mixture of culture medium is kept undisturbed for a long period. The heavier cellular debris settles at the bottom. The culture medium with the recombinant protein becomes the supernatant.
Electroporation is a technique used for introduction of foreign DNA into bacterial or plant cells by subjecting them to a very high voltage or electrical pulse. This results in creation of transient pores in the cell membrane through which the foreign DNA can easily enter the cells.
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