Antibodies play a substantial role in modern medicine, from diagnostics to therapeutics. The immune system produces these specialized proteins to identify and neutralize foreign substances (antigens), like bacteria, viruses, cancer cells, etc. B lymphocytes manufacture antibodies and circulate them throughout the body, attaching themselves to their specific antigens and removing them from circulation.
Antibody production traditionally involved immunizing animals and harvesting the antibodies from their blood. However, this technique possessed several risks that discouraged researchers and scientists from employing it. These risks include varying antibody quality, animal suffering, and difficulty producing human antibodies.
Fortunately, biotechnology has revolutionized how we produce antibodies. Biotechnology entails using living organisms, cells, or their components to generate new processes or products. Scientists can maximize biotechnology to produce antibodies efficiently and without many risks.
This post will discuss the role of biotechnology in shaping antibody production. We will also explore the key techniques and technologies in biotechnology-based antibody production and their benefits over conventional methods. Please continue reading to understand how this technology can accelerate antibody production and increase the chances of succeeding in experimental research.
What is Antibody Production?
Before we define the term “antibody production,” it’s important to understand that it has specific and general meanings. Generally, antibody production refers to developing usable specific antibodies, including preparing the immunogen, immunizing it, creating hybridoma, and collecting, isotyping, screening, purifying, and labeling it for use in a certain technique. In a restricted sense, antibody production refers to all the steps that lead to antibody production except the various forms of labeling and purifying antibodies for specific uses.
Moreover, antibody production entails preparing antigen samples and injecting them into farm or lab animals safely to induce high expression levels of antigen-specific antibodies in the host’s serum. These antibodies are then collected from the animal for further research and applications.
Polyclonal antibodies are collected straight from the serum. On the other hand, monoclonal antibodies are generated by fusing spleen cells that secrete antibodies from immunized hosts with immortal myeloma cells to produce monoclonal hybridoma cell lines. These cell lines express themselves in the specific antibody through cell culture supernatant.
Having said that, the production process can only be successful if you ensure thorough planning and implementation. Here are some critical steps involved in the process:
Purify or synthesize the target antigen (hapten or peptide)
Select the right immunogenic carrier protein
Conjugate the carrier protein and antigen to establish the immunogen
Immunize the host using the right adjuvant formula and schedule
Screen the hybridoma (serum) for antibody isotype and titer (also called antibody characterization)
Biotechnology Technologies Used in Antibody Production
Biotechnology depends on living organisms and their parts to design new scientific processes and products. It employs advanced methods to develop potent antibodies designed for specific targets. Let’s discuss some of the fundamental techniques involved in antibody production:
Recombinant DNA technology
Researchers and scientists often use recombinant DNA technology to insert genes that code specific antibodies into the host cell, like yeast or bacteria. Consequently, the host cell generates the desired antibodies in massive quantities, facilitating cost-friendly and effective production. This technology has transfigured antibody production by creating completely human antibodies, eliminating the danger of hostile immune reactions.
Recombinant antibodies are generated in expression systems that mammalian cell lines develop instead of E. coli cells. The production process begins by isolating the desired nucleic acids (genetic material) or using a gene library with random antigen-binding sequences and then using them in antibody engineering.
The desired genes are administered in expression vectors using the antibody phage display technique. The bacteriophage library is then exposed to immobilized antigens, and strong binders are separated from weak ones, which attach to the antigen. Repeating this selection process with stringent conditions results in the library’s most specific and potent antibodies. As a result, manipulating these genes forms new antibodies with reduced immunogenicity.
Hybridoma technology
Hybridoma technology was developed by Cesar Milstein and George Kohler in 1975. It entails fusing cancer cells with normal antibody-generating cells, creating immortal B cells that release single, specific antibodies. This method is instrumental in making highly-specific monoclonal antibodies suitable for therapeutic applications and assay development. Furthermore, the mammalian origin of these cells can integrate into in vivo post-translational adjustments, reducing the risk of recognition or aggregation failures.
However, the hybridoma technique has a few significant drawbacks worth noting. First, the development process takes a long time (around 6 to 8 months) to attain reasonable antibodies. Second, since the resulting antibodies have a murine origin, they must be humanized to suit therapeutic purposes, which could incur additional charges. These challenges have led to hybridoma antibody production being replaced progressively by quicker, cheaper, and more effective technologies for bio-therapeutic development.
Phage display technology
Phage display is the alternative to the hybridoma technique for drug and therapy development. It was created in 1985 by Smith and involved integrating a gene sequence coding of a certain antibody into a filamentous bacteriophage’s DNA sequence. This integration facilitates the expression on the bacteriophage capsid’s surface and establishes the connection between the phenotype and genotype.
The phage introduces Escherichia coli and utilizes its replication system to display new phages consistently without killing the host cell. As a result, the desired antibodies are produced faster and in large numbers. Consequently, a library of immune or nave phage is constituted, and researchers use it to identify the desired antigen-antibody interactions using screening techniques.
Furthermore, libraries can be developed from any animal, including human beings, making it possible to screen human antibodies directly. Accessing the sequence is relatively easy since the phenotype is linked to the genotype, promoting further recombinant protein production.
The difference between phage display and hybridoma techniques is that the former is fast and lasts a few weeks. It also allows the researcher or scientist to screen various antibodies. Therefore, anti-venom and toxicology research often employ phage display since it can work with non-immunogenic and toxic antigens.
The Bottom Line
Antibodies are fundamental molecular sentinels highly specific for their respective antigens, and incorporating biotechnology in this field has contributed immensely to therapeutics and life science research. Traditional techniques generate antibodies via immunization, creating antibodies with different affinities and specificities. On the other hand, biotechnology produces antibodies that specifically identify and attach to one target with high affinity.
Biotechnology also allows scientists to produce human antibodies that are less likely to produce harsh reactions, making it ideal for sensitive therapies and scientific solutions. Therefore, biotechnology has been crucial to the success of antibody production and will continue doing so in the future.
The post Revolutionizing Antibody Production – The Role of Biotechnology appeared first on Datafloq.
