Antibodies as proteins are quite fascinating. They are a significant part of your body’s immune response in which these proteins are naturally produced to fight viruses that enter the body. They are also protective agents that prevent the development of diseases like cancer.
Apart from these vital functions, antibodies are also unique in their high specificity and affinity. This property allows them to be one of the most crucial research tools in biochemistry and other fields of science or medicine.
Due to the roles recombinant antibodies play in research, their production in the laboratory is equally valuable. This article will demonstrate some key things you need to know to produce these important biological markers successfully.
What Are Recombinant Antibodies?
Recombinant antibodies (rAbs) are monoclonal antibodies that are produced in vitro through the use of synthetic genes. Contrary to monoclonal antibodies (mAbs) generated by traditional hybridoma-based technologies, you don’t require hybridomas or animals during recombinant antibody production.
These antibodies and their monoclonal counterparts can serve different purposes in toxicological research and biomedical science. They can also be vital as effective treatments for autoimmune disorders, cancer, and other medical conditions. But though the use of monoclonal antibodies has become widespread in biomedical science and medicine as a result of their ability to attach and destroy or neutralize cell-specific antigens, their production through the ascites method puts animals used in the process through immense pain.
Consequently, the governments of countries such as the United Kingdom, Australia, the Netherlands, and Germany prohibited this method in favor of in vitro production. In the U.S, in vitro method is regarded as the standard technique for mass production.
Recombinant Antibody Production
The production of recombinant antibodies begins with isolating nucleic acids or genetic material coding for antibody subjects. In some cases, a library of genes also serves the same purpose: to have randomized antigen-binding site sequences for antibody production.
These genes are placed in expression vectors that show the associated antibodies on their surface. This method is known as the antibody phage library technique. In another process called the panning technique, a bacteriophage library is introduced to an immobilized antigen; the weak and robust binders can be separated by washing. The stronger binders bind to the antigen if special agents are not used.
This selection process undergoes numerous repetitions under increasingly rigorous conditions, which leaves in the antibody library only the most specific and strongest antibodies. This in vitro enables the producer to create new antibodies by manipulating the genes. It also limits immunogenicity or only applies to antibody fragments like fab or scfv fragments.
Then, the most viable antibody genes undergo cloning into the appropriate cell lines that serve as expression platforms. Due to the relatively complex nature of antibody proteins and their tendency to undergo modification after expression, eukaryotic or higher cells are typically required.
This means bacteria cells such as escherichia coli would be incapable of producing the antibodies desired. Yeast cells are better suited. But Chinese hamster ovary (CHO) cells and human embryonic kidney (HEK) cells are the best options that yield high-quality antibodies.
Who Can Produce Recombinant Antibodies
Specialists in the production of recombinant antibodies provide the required capacity and expertise for qualitative and rapid antibody expression. As such, the majority of the companies or organizations that use recombinant antibodies outsource their production to more specialized companies.
To create a monoclonal antibody, research needs to start by identifying the precise antigen to attack. Among the vital factors that researchers need to consider for function antibodies are:
- Immune response
- Effector Functions
- Long-term efficacy
During cancer therapy, the process needs the isolation and characterization of rAbs targeted toward the specific tumor cell. Owing to the results obtained from research methodology and current mammalian cell culture optimization, the production of human antibodies in these expression systems could be enhanced to more than 12 g/liter, without the need for animals.
Advantages of Producing Recombinant Antibodies
The ability of recombinant antibody technology to circumnavigate the several drawbacks of conventional monoclonal antibody expression cannot go unmentioned. At the same time, it also offers multiple inherent advantages and a much broader scope of application. Here are some of the more remarkable benefits of recombinant antibodies.
- Best quality – Since optimizing the desired genetic material is easy, rAbs are designed to have superior sensitivity, specificity, and affinity.
- Scalability – The process of incorporating an established and optimized genetic sequence into host cells is relatively easy.
- Highly consistent production – rAbs production lacks the spontaneous mutations that are common in hybridoma cell lines.
- Diverse antibody production – The technology is agile enough to enable the rapid expression of different antibodies.
According to some, recombinant antibodies embody the present and future of antibody production and research. Their numerous advantages over monoclonal and polyclonal antibodies make it easy to see why recombinant Abs are favorable tools with bright futures for clinical and research purposes. Some of these advantages, like complete control over their production, the ability to engineer, the possibility of Ab/antigen binding, and the lack of animal subjects, contribute to the increasing popularity of rAbs for clinical, research, and diagnostic applications. At present, many commercial antibody providers have begun shifting their hybrid-generated monoclonal Abs to rAbs.