Human insulin, an indispensable molecule for the body’s metabolism, is fascinatingly similar in structure to the insulin produced naturally by our pancreas. How is this possible? How is it that we can replicate a crucial hormone so precisely? In the heart of the process lies the answer: Recombinant-DNA technology and a small component called a homogenizer.
Understanding Insulin and Its Structural Identity
Insulin, a hormone produced by the pancreas, plays a critical role in the body by allowing cells to take in glucose and use it for energy. When we say “human insulin,” we are referring to insulin that is structurally identical to what our body naturally produces.
The Role of Recombinant-DNA Technology
This structural identity is achieved through the use of recombinant-DNA technology. Sounds technical, right? Just think of it as a method that allows scientists to create DNA sequences that result in the desired insulin.
The Use of Production Organisms
What are the actors of this grand performance? Two key players are Saccharomyces Cerevisiae (baker’s yeast) and E. Coli, used as production organisms. They aid in the production process by contributing their cellular machinery to produce the human insulin.
The Insulin Production Process
The journey from DNA sequence to human insulin is an intricate one.
Fermentation of Cells
It begins with the fermentation of cells. Here, cells grow and multiply in a controlled environment, producing the proteins that will eventually form insulin.
High Pressure Homogenization
Then, we arrive at a critical stage, high pressure homogenization. This process disrupts the cells, releasing the proteins. Imagine a speed centrifuge spinning rapidly to concentrate cells. These cells are then resuspended with a buffer to prepare for the high-pressure homogenization.
Advantages of High Pressure Homogenization
There are several advantages to high-pressure homogenization. It’s quick, efficient, and replicable on an industrial scale. Moreover, it’s a mechanical process that doesn’t require the addition of expensive chemicals or enzymes.
Cell Disruption and Protein Extraction
This process of cell disruption releases the intracellular material, which, post cooling and centrifugation, gets separated from the cell debris. The efficiency of this process can be evaluated by the content collected after the centrifugation. The proteins are then processed further to construct the peptide chains characteristic of human insulin.
Delving into the Homogenization Process
In the homogenization process, the product is kept at low temperatures (below 10°C) and is homogenized at a high pressure of 1000 bar.
The Impact of Multiple Passages
To achieve efficient cell disruption, more than one passage through the homogenizer is usually required.
The Effects of Homogenization on Insulin Production
Homogenization plays a crucial role in insulin production.
Role in Cell Disruption
Insulin, being an intracellular material, requires cell disruption for its extraction. High-pressure homogenization proves to be an excellent method for large-scale cell disruption.
Importance of Complete Cell Disruption
Complete cell disruption is crucial because any undisrupted cells will co-sediment with the intracellular material in the subsequent solid-liquid centrifugal separation.
The Ideal Homogenizing Machine
To effectively carry out this process, a homogenizer designed for high pressures between 1000-1500 bar is used.
Design Features for Insulin Production
This homogenizing machine has a VHP compression block for high abrasion and high viscosity, with a knife-edge design valve ideal for cell disruption.
Conclusion
The journey of producing human insulin is a testament to the marvels of science and technology. It is a meticulous process that involves the mastery of several steps, with homogenization playing a pivotal role. The process of high-pressure homogenization, the mechanism of cell disruption, and the extraction of intracellular proteins all come together to produce insulin, structurally identical to that produced by our pancreas.
FAQs
- The homogenizer plays a crucial role in disrupting cells to release the proteins that form insulin.
- High-pressure homogenization is a quick, efficient, and replicable process that doesn't require the addition of expensive chemicals or enzymes.
- Saccharomyces Cerevisiae (baker's yeast) and E. Coli are used as production organisms in the insulin production process.
- The efficiency of cell disruption can be evaluated by the content collected after the centrifugation.
- Complete cell disruption ensures that all the necessary proteins are extracted, and no undisrupted cells interfere with the subsequent solid-liquid centrifugal separation.
