Table of Contents:
- Introduction
- What are Recombinant Proteins?
- The Production Process of Recombinant Proteins
- Applications of Recombinant Proteins
- People Also Ask | FAQs | Summary
- IBI Scientific: Your Trusted Recombinant Protein Solution
Introduction
When you think of recombinant proteins, what comes to your mind?
Perhaps terms such as cloning and genetic engineering peak your interest. Maybe its the prokaryotic or eukaryotic systems that make it easy to manipulate genes. In this comprehensive blog, we will explore the ins an outs of recombinant proteins.
What are Recombinant Proteins?
Recombinant proteins are proteins that are engineered genetically usually in a lab. These specialized proteins are made by putting a gene into a protein, and then putting that protein into an expression system. Expression systems can vary, but normally its bacteria, yeast or sometimes mammalian cells. By using said expression systems, proteins can be created on a mass scale allowing for easy duplication and purification.
The Production Process of Recombinant Proteins
1. Gene Cloning
Gene cloning is the very first step in the production process of recombinant proteins. It consists of isolating the gene that encodes the protein and inserts it into appropriate carriers, like a plasmid for example. The carrier is an essential vehicle that presents the gene to the host cells for transformation.
2. Transformation
Once the gene is put into a vector, the carrier can then be put into the host cells. This is called transformation, and it can be accomplished through a pleothra of different methods. Methods such as heat shock or chemical treatment can transform the host cells. As a result, the host cells will start holding on to recombinant protein DNA, being able to express the protein.
3. Expression
After this, the host cells will be grown under optimal conditions to support the expression of the recombinant protein. The expression host can range from bacteria, yeast, insect, and mammalian cells based on the post-translational modifications and the overall yield requirements needed for the protein.
4. Purification
Following the expression of the recombinant protein in host cells, purification of this expressed product is essential. Purification techniques used include chromatography techniques, usually affinity, ion-exchange, or size-exclusion chromatography. Purification ensures a high-purity, contaminant-free final product.
5. Characterization
Characterization is a process that confirms the protein's identity, purity, and biological activity. Amino acid sequencing and MS are techniques often used to certify identity. Amino acid sequencing gives information on the sequence of the protein with atomic resolution. MS provides very accurate molecular weight and post-translational modification information. The key verification technique is Western blotting, where proteins are separated by SDS-PAGE and then transferred onto membranes, which are probed with specific antibodies for identification and size confirmation of the target protein.
Purity assessment uses several techniques to prove the absence of contaminants in the recombinant protein. SDS-PAGE is an important separation technique by molecular weight, and a single sharp band indicated high purity. HPLC allows separation of the components of a protein mixture to be quantified, providing full information about purity and homogeneity. Size-exclusion chromatography, SEC, or gel filtration, separates proteins on the basis of size and shape, thus detecting aggregates and making sure that the monomeric form of the protein is the most prevalent. All these methods ensure that the recombinant protein to be used for further applications in research, medicine, or industry is pure, correctly folded, and biologically active.
Amino acid sequencing gives information on proteins.
Applications of Recombinant Proteins
1. Medicine
Recombinant proteins in medicine are associated with the development of therapeutic agents, vaccines, and diagnostic tools. One important application is the production of insulin using recombinant DNA technology crucial for the treatment of diabetes. Other examples of recombinant proteins that have been very vital in the treatment of a number of diseases and disorders are the erythropoietin, growth hormones, and monoclonal antibodies.
2. Research
One of the very key tools in scientific research includes recombinant proteins. They help to study protein structure and function, investigation of the cellular processes, and developing new assays and techniques. Among the most commonly used recombinant proteins is the green fluorescent protein, which makes the examination and tracing of cellular processes in real time possible.
3. Biotechnology
In biotechnology, recombinant proteins are used to manufacture enzymes, biofuels, and industrial chemicals. Application of enzymes obtained with the help of recombinant DNA technology is realized in many industries, for example in food, beverages, pharmaceuticals, and agriculture. Most often, such enzymes have improved stability, specificity, and efficiency than their natural counterparts.
4. Agriculture
Such recombinant proteins in agriculture help in better crop yields, resistance to pests and pathogens, and GMOs. Examples include recombinant bovine somatotropin, rBST, which acts to increase milk yield in dairy cattle, and recombinant vaccines protect from infectious diseases in livestock.
People Also Ask | FAQs | Summary
What are examples of recombinant proteins?
Recombinant proteins are diversified and have huge applications in many areas. Some examples include insulin, which is used against diabetes, and monoclonal antibodies like trastuzumab (Herceptin) applied for cancer treatment. Other remarkable examples are: erythropoietin that stimulates the production of red blood cells, and green fluorescent protein (GFP), a tool in molecular biology to study gene expression and protein localization.
Why are recombinant proteins useful?
Such recombinant proteins become highly useful for therapeutical purposes, as well as in the realm of scientific research or industrial processes, due to the large quantity producible with high purity and consistency. This would enable the production of proteins that otherwise may be difficult to isolate from natural sources. That would mean such things as drugs, vaccines, and enzymes could be more suitably and cost-effectively made.
What are recombinant proteins for medical use?
In the medical field, recombinant proteins are used as therapeutic agents, vaccines, and diagnostic tools. Examples of recombinant proteins include insulin for diabetes, growth hormones for growth disorders, and clotting factors for hemophilia. Other critical medical applications include monoclonal antibodies, such as those used in cancer treatment, and recombinant vaccines, such as that against hepatitis B.
Why is recombinant protein called so?
It is called recombinant protein because it is a product expressed through recombinant DNA technology. Basically, what this means is that a gene is inserted into a host organism's DNA; from then on, the host will be able to express and produce the protein. The term "recombinant" literally means that DNA from different sources is combined in order for the target protein to be expressed.
How does protein differ from a recombinant protein?
The main difference between a natural protein and a recombinant protein is in the method of its production. The former are proteins produced by living organisms as part of their normal biological activity. In contrast, the latter are genetically engineered through the insertion of a gene of interest into a host organism, which gets expressed and then produced in the laboratory.
How do scientists produce recombinant proteins?
These researchers working on this isolate the gene coding for that particular protein and clone it into a suitable vector. Such a vector is then transferred to host organisms, either bacteria, yeast, or even mammalian cells, via a process known as transformation or transfection. Such host cells are induced to grow under the best possible conditions, followed by the expression of proteins. The expressed proteins are finally purified to high purity and activity using techniques such as affinity chromatography.
IBI Scientific: Your Trusted Recombinant Protein Solution
Recombinant proteins in modern biotechnology have become the cornerstone between medicine and agriculture. The complexity of their creation involves sophisticated techniques for genetic engineering, expression, and purification; each is important for obtaining a high-quality, functional protein. IBI Scientific provides tools and reagents to help give your recombinant protein production the best opportunity for success. We have many products including: purification kits, reagents and proteins that will help raise your research and development projects to new heights. Whether you are doing simple or even advanced research, IBI Scientific is dedicated to helping you in the interesting world of recombinant proteins.
