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Antibodies have been a critical reagent in biological research for decades. Recently, due to issues surrounding scientific reproducibility, there is a growing need for antibodies with extraordinary specificity and minimal lot variability. Until recently, this was not possible. However, recent advances in molecular biology and screening have enabled production of genetically defined, recombinant antibodies.
Due to their low lot-to-lot variability and high specificity, recombinant antibodies are being touted as the future of antibodies in research paving the way for researchers to have unrivalled reproducibility and trust in these reagents.
There are many methods of producing recombinant antibodies. At Proteintech, recombinant antibodies are monoclonal antibodies that have been produced by in vitro cloning of the antibody heavy and light chain DNA sequences from the B cells or plasma cells of immunized animals. Unlike traditional monoclonal antibodies produced from hybridomas, the recombinant vectors are introduced into expression hosts (e.g. E. coli) to then produce the antibodies. This recombinant technology results in almost no lot-to-lot variability, removing the risk of genetic drift that can result in variations in of monoclonal antibodies.
The storage of DNA libraries of antibody sequences and subsequent use of recombinant technology ensures that antibody production is highly consistent and reliable, minimizing the risk of genetic drift and providing superior antibody supply over time.
Due to the nature of recombinant antibody production (link to section), recombinant antibodies have a decreased production time and large-scale manufacturing of recombinant antibodies is easier to achieve. Therefore, with almost no variation between batches and the ease of scalability, recombinant antibodies are perfect for long-term projects or experiments where high volumes of antibody are required. Monoclonal and polyclonal antibody production can sometimes face issues of low yield, therefore using recombinant production enables manufacturers to circumvent this issue.
Once the sequence of an antibody has been cloned for recombinant production, it is then possible to modify the sequence in order to generate antibodies which can bind with higher affinity and specificity to its target. Antibody engineering also enables the creation of antibody “chimeras”, whereby the Fc portion of the antibody can be changed to that of a different species. (see CTK chimera section).
Some targets, such as specific protein isoforms or phosphorylated proteins can be very difficult to produce high quality and reliable antibodies using mouse hybridoma technology. Due to recombinant technology enabling antibody engineering from species with richer immune diversity than mice like rabbits and llamas,specific antibodies towards these hard targets can be produced.
The need for animal-free technologies is increasingly required and encouraged around the world. In recombinant production, once the antibody sequence is cloned following immunization, the need for animals is eliminated.This process can even create antibodies from non-traditional sources. For example, our SARS-COV-2 S-protein recombinant antibodies (Link to COVID-19 page) are derived from small samples of convalescent patient sera.
At Proteintech, we create rabbit recombinant antibodies. Rabbits have superior qualities for recombinant antibody production over other methods, harnessing their natural immune response for its evolutionarily honed epitope recognition, high antibody diversity, and single isotype subclass.
Proteintech’s recombinant antibody development starts with rabbit immunization. As the immune response peaks, antigen-specific single B cells from the rabbits are isolated using our proprietary technology. The selected B cells are then screened for their ability to produce antibodies with highest sensitivity and specificity. The genes for the antibody heavy and light chains from the selected B cells are then cloned into cell lines to express the desired antibody. These antibodies are extensively validated, adhering to the IWGAV (International Working Group on Antibody Validation) recommendation to prove the specificity, sensitivity, and reproducibility.
Nanobodies, the binding domains of heavy chain antibodies from Camelidae family members, can also be fused to Fc-domains to generate chimeric heavy chain antibodies. The Fc-domain fusion format combines the advantages of Nanobodies with those of traditional antibodies: The VHH/Fc fusions (i) are bivalent so they have even higher avidities and affinities than the parent Nanobodies, (ii) bind unique, 3-dimensinal conformations of epitopes not recognized by traditional antibodies, and (iii) can be detected and captured using tools from the broad range of antibody reagents like secondary antibodies and Nano-Secondaries. Like other recombinant antibodies, VHH/Fc-fusions are available with high lot-to-lot consistency at an unlimited supply. The Nanobody Fc-fusion recombinant antibodies have been developed in collaboration with Absolute Antibody Ltd.
Proteintech and ChromoTek offer nanobodies fused to two different Fc-domains - rabbit IgG and mouse IgG1 – for flexibility in your experiments. The popular ChromoTek Nanobodies anti-GFP-, anti-mNeonGreen, anti-TurboGFP-, and anti-vimentin-VHHs are all available in chimeric rabbit IgG and mouse IgG1 form.
|GFP VHH-Fc fusion|
|GFP recombinant antibody, VHH-mouse IgG1 Fc fusion [CTK0201]||gfms||IF||mouse IgG1 Fc|
|GFP recombinant antibody, VHH-rabbit IgG Fc fusion [CTK0201]||gfrb||IF||rabbit IgG Fc|
|mNeonGreen VHH-Fc fusion|
|mNeonGreen recombinant antibody, VHH-mouse IgG1 Fc fusion [CTK0203]||nfms||IF||mouse IgG1 Fc/td>|
|mNeonGreen recombinant antibody, VHH-rabbit IgG Fc fusion [CTK0203]||nfrb||IF||rabbit IgG Fc|
|TurboGFP VHH-Fc fusion|
|TurboGFP recombinant antibody, VHH-mouse IgG1 Fc fusion [CTK0204]||tbfms||IF||mouse IgG1 Fc|
|TurboGFP recombinant antibody, VHH-rabbit IgG Fc fusion [CTK0204]||tbfrb||IF||rabbit IgG Fc|
|Vimentin VHH-Fc fusion|
|Vimentin recombinant antibody, VHH-mouse IgG1 Fc fusion [CTK0211]||vfms||IF||mouse IgG1 Fc|
|VVimentin recombinant antibody, VHH-rabbit IgG Fc fusion [CTK0211]||vfrb||IF||rabbit IgG Fc|