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NCERT Exemplar · Q16

Q.Which of the following should be chosen for best yield if one were to produce a recombinant protein in large amounts?

(a) Laboratory flask of largest capacity
(b) A stirred-tank bioreactor without in-lets and out-lets
(c) A continuous culture system
(d) Any of the above
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To produce recombinant proteins in large quantities, a continuous culture system is the most efficient choice because it maintains cells in their most productive exponential growth phase.

When we talk about producing recombinant proteins in large amounts, we are moving beyond the small-scale experiments of a laboratory bench and into industrial biotechnology. The goal is to maximize the yield of the desired product, which means creating the most favorable conditions for the host cells (like bacteria or yeast) to grow and synthesize the protein.

Initially, experiments might be done in small laboratory flasks. While useful for initial studies, these flasks are inherently limited in volume and control. As the cell density increases, nutrients are quickly depleted, and metabolic waste products accumulate, leading to a decline in cell growth and protein production. This is a classic example of a batch culture system, where the entire process runs from start to finish without adding fresh medium or removing spent medium.

To overcome these limitations and achieve large-scale production, specialized vessels called bioreactors are employed. Bioreactors are essentially large containers designed to provide an optimal environment for microbial or cell culture. They are engineered to control various parameters crucial for cell growth and product formation, such as:

  • Temperature: Maintained at the ideal level for the specific organism.
  • pH: Kept stable using acid/base addition systems.
  • Substrate (nutrients): Supplied adequately for cell metabolism.
  • Salts and Vitamins: Essential micronutrients are provided.
  • Oxygen: Crucial for aerobic processes, often supplied through sparging and agitation.

Among the common types, stirred-tank bioreactors are widely used. They feature an agitator system for uniform mixing of contents, an oxygen delivery system, a foam control system, and systems for temperature and pH control. The stirring ensures that oxygen is available throughout the culture and that nutrients are evenly distributed.

Note

The primary purpose of a bioreactor is to provide an environment where cells can grow optimally and produce the desired product in high concentrations. Without proper control over growth conditions, even a large vessel will not yield good results.

Now, let's consider the different operational modes for bioreactors, which directly impact the yield.

  • Batch Culture: This is the simplest mode, similar to a laboratory flask but on a larger scale. The bioreactor is inoculated with cells, and all nutrients are added at the beginning. The culture proceeds until nutrients are exhausted or waste products become inhibitory. Cell growth follows a typical curve: lag phase, exponential (log) phase, stationary phase, and death phase. Protein production is highest during the exponential phase, but the overall yield is limited by the eventual decline in cell activity.

  • Continuous Culture System: This is a more advanced and efficient method for large-scale production. In a continuous culture system, fresh culture medium is continuously added to the bioreactor from one side, while an equal volume of the spent medium (containing cells and products) is simultaneously drained from the other side. …

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