Recombinant antibodies, how are they made and what are the benefits versus other animal models?

Overview:

Recombinant antibodies (rAbs) are monoclonal antibodies produced in vitro by using synthetic genes. The main difference between them and monoclonal antibodies is that with the arrival of the recombinant antibodies several significant drawbacks associated with mAbs are no longer an issue you have to deal with in your experiments. To better understand how they are generated, what are their benefits and why they might just be the better option for your experiments, please read on.

• What are Recombinant antibodies(rAbs) and how are they produced?
• But do they overcome the drawbacks of using classical monoclonal antibodies?
• So, should you consider using them?

What are Recombinant antibodies(rAbs) and how are they produced?

This type of antibodies is generated in vitro using synthetic genes outside the limitations of the immune system. The process begins with isolating antibody genes from source cells, multiplying and cloning them in a suitable phage vector and then introducing the vector inside a host: mammalian cell line, bacteria or yeast. These steps result in adequate amounts of usable antibody. The produced rAbs can be used in all the applications, usually associated with classical monoclonal antibodies.

Furthermore, recombinant antibodies can be cloned from all antibody-producing animal species if suitable oligonucleotide primers or hybridization probes are present. The power of manipulating the antibody genes greatly contributes to synthesising new antibodies and fragments in vitro. Creating display libraries in phage or yeast gives us the option of analysing and selecting the suitable characteristics emerging from the changes in antibody sequence.

With all the unlimited possibilities and types in the display libraries, a question arises: What is the easiest and most efficient way of choosing the ones that display the antibody needed? This can be easily achieved by using a procedure known as panning. It is a selection process against the desired antigen. The easiest way to perform panning is to use a variation of a well-known ELISA procedure. First, the antibody must be incubated alongside the target immobilised on a solid phase, an ELISA plate in this case. Second, washing removes the unbound phage and specifically bound phages are eluted and amplified by infecting E. Coli cells. This step is repeated three to four times in order to isolate the phages that display antibodies with the highest stability and affinity. After that, genes for the desired antibodies are sequenced and if needed, they can be subjected to affinity maturation. Lastly, the genes related to the best antibodies are transferred in a suitable expression system and are ready for mass production.

But do they overcome the drawbacks of using classical monoclonal antibodies?

Recombinant antibodies have several significant advantages when compared to mAbs.

First, they have much higher reproducibility and control, mainly because recombinant production offers more control over the antigen, mainly because rAbs are defined by the sequence responsible for their encoding, making them the more reliable option of the two.

Second of all, the time needed to produce an antigen specific antibody can be brought down to eight weeks in some cases, compared to at least four months needed if hybridoma technology is used.

Also, a recombinant antibody fragment of interest can be easily converted in different isotype, subtype or species, just by adding the suitable constant domain, helping with switching antibodies into a more user-friendly format.

Finally, an advantage, that has been overlooked: Some types of rAbs can be synthesised without the use of any animals in the process, thus eliminating the animal welfare and ethical concerns associated with classical mAbs production. Regarding the pain and discomfort inflicted on animals during the production of monoclonal antibodies, several countries, including Germany, the Netherlands, Australia and the UK have banned their production in favour of new in vitro methods. However, in vitro methods with hybridomas have several limitations, including immunization and euthanasia of the animals, being slow and intensive and they often must be humanised before being administrated to patients, in order to avoid immune reactions.

So, should you consider using them?

Generally speaking, yes, you should consider using them. They can be constructed against any antigen, even human and highly conserved ones, which are not recognised as foreign by the mouse’s immune system. The process of humanisation is not needed for rAbs, mainly because they circumvent our immune system response which is elicited by murine mAbs. Also, they can fuse with toxins and drugs, making them usable in therapeutics. The less time required for their production, impressively flexible selection process and being much more reliable and having greater reproducible results compared to classical monoclonal antibodies makes rAbs the better option in most of the cases.

In order to help our customers with their choice of antibody with low risk, we offer one of the largest ranges of primary antibodies with trial size options available.

The Antibody Validation Project works like this.
1. Order your trial size antibody online.
2. Explore its suitability.
3. Return your results by posting them on the product page.
4. Receive a sample refund.

Find out more here.