Rabbit monoclonal antibody service (recombinant discovery)

Boster Bio PCD custom rabbit monoclonal antibodies (Recommended)

Best Choice For Diagnostic And Therapeutic Antibodies

Recombinant rabbit monoclonal antibody production features high affinity, easy humanization and high scalability. The pricing is more than what one would expect for developing research only but it is very affordable for diagnostic and clinical antibody discovery projects. The turnaround time is around 6 months.

As part of the humoral immune system, B-cells normally begin as stem cells in the bone marrow (in mammals), or the bursa of Fabricius (in birds), and then migrate to the spleen. There they undergo somatic mutation with 1% developing into antigen-specific plasma cells which secrete large amounts of antibodies.

Therefore, starting with the process of isolating antigen-specific plasma cells reduces screening time and ensures a larger selection of antibodies with the highest antigen affinity, making Boster’s PCD technology most efficient way to isolate high-affinity antibodies.

Boster’s PCD technology employs a two-step screening procedure; a large scale efficient Fluorescence activated cell sorting (FACS) to isolate antigen-specific plasma cells from millions of splenocytes followed by secondary testing of individual plasma cells for their recombinantly expressed antibodies in the supernatant. By isolating antigen-specific plasma cells on FACS, PCD technology increases 500-fold the number of antibody clones that can be screened. Therefore, the ability to find the best antibodies, which may be rare and difficult to find with traditional methods, is greatly enhanced.

Hybridoma Rabbit Monoclonal

First Generation RabMab Technology

Even though it shares the same same CDR repertoire, hybridoma rabbit monoclonal technology falls short on the screening scale. The last generation rabbit monoclonal technique only screen thousands of average splenocytes that were lucky enough to successfully fuse while as we can screen the whole spleen if we want to. Our workflow does not require to keep the cells alive while screening and we go for the cream of the crop only -- plasma cells, with the brightest signals. Some other disadvantages of the traditional Hybridoma Rabbit Monoclonal technology:

• >99% of cells may not complete or survive the fusion process
• Cultures prone to contamination
• Poor recognition of non-immunodominant epitopes
• ‘one shot’ analysis – non-fused products cannot be recovered if initial analysis unsuccessful
• Low yield of conversion from target clones to IgG
• Origin plasma cells express high immunogenicity
• Not well developed for systems beside rabbit and mouse

Mouse Monoclonal

Had Its Days, Still Good For Research

Mice have limited CDR repertoires. In general mammals with longer reproductive cycles tend to have larger CDR repertoires due to the evolutionary pressure of survival long enough to reproduce. Mice excel in their innate immunity but their adaptive immunity is not as robust as that of rabbits. This puts mice at a disadvantage for generating antibodies with high affinities.

Rabbit Polyclonal

A Quick And Cheap Way To Make Some Antibodies

Long story short, polyclonal antibodies work for research but not for diagnostics. It is indeed a very low cost and fast turn around method for making antibodies against certain targets. Boster Bio does offer a $600 package for making a polyclonal antibody. Some claim that polyclonal antibodies suffer from specificity because they originate from multiple clones. This is a misconception because this "disadvantage" can be mitigated by using a single epitope for immunization and purification of the polyclonal antibodies. There are 3 main disadvantages of polyclonal antibodies in our experience. First is lot-to-lot variability. For any clinical assays or therapeutic assays it is almost impossible to get approval for an assay with polyclonal antibodies as core components. For research use however it is acceptable, and is often used as a stepping stone towards a monoclonal antibody. In our experience most antigens that prove to generate a good polyclonal antibody can also make a good monoclonal antibody. The second disadvantage is its inability to amplify only the best clones. This is important if the target protein is difficult to detect and requires high affinity. This is often the case in detecting secretive proteins and biomarkers in liquid biopsies, via ELISA or other similar immunoassays. During affinity chromatography, all antibodies with enough affinity to bind to the very concentrated antigens will be pulled out. Many of these antibodies might not have enough affinity to bind to the target protein in vivo where the target protein exists in low levels and masked by competing binding receptors in the matrix/in vivo environment.

Phage Display

2nd Best Choice For Diagnostic And Therapeutic Antibodies

It lacks SOMATIC HYPERMUTATION. Even though phage display allows a reasonably large screening scale, it still falls short when compared to screening the full spleen, by at least a 1000 fold in scale. But the real problem is the CDR library phage display platforms have are "diamonds in the rough". Without going through SOMATIC HYPERMUTATION they will not be able to bind to the intended antigen with high affinity. Even though some claim they have secrete sauces in antibody engineering to improve antibody affinity after the fact, studies and our experience indicate that it is often no match to mother nature's affinity maturation process. It is a numbers game after all and the immune system can go through billions of iterations in vivo in a matter of weeks while as antibody engineering needs to experiment one amino-acid modification at a time, at weeks per iteration. Edited: some customers mentioned that one can train the phage library with certain antigen/pathogen groups to have better results. While this certainly helps, it is not a magic bullet to solve the intrinsic issues with phage display. I would also add that phage display does have an advantage, among many others, that it is great for generating antibodies against antigens that are not immunogenic or are autoimmunogenic, as it circumvents negative selection. See more information here Central Tolerance Wikipedia.

Timeline and costs