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With our patented microfluidics plasma cell discovery (PCD) platform, you can get fully validated, affinity matured rabbit monoclonal antibodies:
The scale of phage display meets the power of in vivo affinity maturation--antibody discovery has never been so robust.
Difficult targets? No problem! Boster Bio has you covered with rabbit mAbs of the highest affinity and specificity tailored for your desired functionality.
The PCD platform produces the highest affinity/specificity clones amongst all antibody discovery technologies and is best suited for Diagnostics & Therapeutics targets.
Boster Bio's plasma-cell discovery (PCD) technology uses flow cytometry to screen individual cells treated with proprietary chemistry. This unique compound keeps B cells from secreting antibodies, retains antibodies on cell membranes. Subsequently the splenocytes are incubated with fluorochrome conjugated antigens and the plasma cells. This enables the screening of the entire splenocyte population. The clones with the highest brightness have the strongest affinity and are isolated for down stream screening and processing.
Suitable targets: small molecules, DNA, peptides, proteins and cell-based antigens. PCD excels at tackling difficult targets and applications, such as antibody pairing, recognising PTM specificity and point mutations.Get a free consultation
Put It In Context Of Scale
PCD platform can screen the entire spleen of a rabbit. This makes a huge difference in terms of finding rare populations of high affinity clones. The key to developing antibodies for difficult antigens is to screen as much and as early as possible. The amount of clones whole spleen screening can cover is 100-1000 times more than phage display, and millions or more times more than transitional methods like hybridoma. The PCD screening technology brings the best chance to find the rare clones with highest affinity for your biomarkers.
We have successfully created supreme affinity antibodies for Therapeutics targets such as Ion channels, small molecules and GPCR’s.
"Novel clinical biomarkers are increasingly more difficult to make. The easy ones have already been discovered and commercialized. Thus in antibody discovery, finding that top 0.1%, even 0.01% highest affinity B-cell population is key to breakthrough."
CJ Xia, VP Sales and Marketing
It is our project concierges' mission to make your project experience as smooth and memorable as possible. They are subject matter experts who are easily accessible around the clock, always happy to help you solve problems, make recommendations and sort through options.
3 core concepts, move your mouse over to see details.
When building an antibody for diagnostic assays or therapeutic candidate, recombinant antibody is the go to choice. It has the following benefits:
The key to discovering highest affinity clones is to screen as many clones as possible as this ensures a greater chance to isolate the best potential antibodies to your target. Boster Bio's microfluidics plasma cell discovery (PCD) platform can do just that by isolating antigen-specific plasma cells from millions of splenocytes followed by secondary testing of individual plasma cells for their recombinantly expressed antibodies in the supernatant. This increases the number of antibody clones that can be screened by 500 fold. Because no fusion step is required, the cells do not need to stay alive. The cells are fixed our PCD platform can screen for as long as you need to. We can cover the entire spleen and have done so in some cases. Typically 7 days of screening is sufficient for generating very high affinity clones.
|Boster Bio PCD Rabbit Monoclonal||Hybridoma Rabbit Monoclonal||Mouse Monoclonal||Rabbit Polyclonal||Phage Display|
|Affinity||+++High||++Hit&miss||+Low||++Hit&miss||++Can be better|
|CDR Pool Size||+++Large||+++Large||+Small||+++Large||+++Large|
|Applications||Diagnostics & Therapeutics||Diagnostics & Therapeutics
||Diagnostics & Research||Research||Diagnostics & Therapeutics|
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.
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:
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.
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.
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.
See the difference Boster Bio’s validated custom rabbit monoclonal antibody production makes in your success generating actionable insights for therapeutic and diagnostic target candidates.
Boster's custom rabbit monoclonal antibody plasma-cell discovery (PCD) screens the immune repertoire of rabbits. Our aim is to produce an antibody that works for your specific application. Our technical staff will guide you through the project, from designing the antigen, immunization, screening strategy, antibody validation, to the proper handling of the plasmid that expresses your antibody. We can also generate antigens for you. Our antigen synthesis service cover both peptides and recombinant proteins. We are capable to express the antigen in E. coli, baculovirus, and mammalian cell lines.
The final deliverable is the sequences of selected positive clones, and plasmids containing the cloned antibody genes. Recombinant antibodies ensure the preservation of genetic information.
|Peptide and Immunization: (2 rabbits)||Titer Data|
|Peptide synthesis 10mg, >90% purity||1 week||Titer Data|
|Peptide conjugation KLH for immunization and BSA-biotin conjugation for probing and screening||1 week||Titer Data|
|Immunization for 2 rabbits||2-3 months||Titer Data|
|Titer ELISA Assay Monitoring anti-sera titer and selecting the best animal for monoclonal|
|First Screening with FACS (> a million B cells)
|Supernatant production 0.25 ml scale, and Secondary screening with ELISA||1 week||ELISA data|
|Customer further screening with supernatant if necessary, choose three best clones.|
|Clone Deconvolution, cloning, expression/ELISA conformation; 30ml scale production, yield~ 0.5mg Ab per clone||3-4 week||Sequences|
What you will receive upon successful completion of above phases.
Contact your project concierge for more information.
Use for research, IVD and therapeutics
Antibody Generated with PCD Technology
Cyclophosphamide (Cyc), is a chemotherapy drug and immune suppressor which causes side-effects such as vomiting, hair loss and bleeding. Doctors monitor cyclophosphamide metabolism in the blood using an antibody-based assay, to adjust the dosage accordingly and minimize side effects. We tested antibodies generated by PCD tech against commercially available mouse monoclonals.
All four randomly selected clones had a greater affinity for Cyclophosphamide than the commercial mouse hybridoma-based standard. Rabbit antibodies have a greater affinity to their antigens than mouse antibodies in general, and the PCD technology adds to this by screening potential candidates for greater efficiency.
Figure 1. Rabbit anti-Cyc monoclonal antibodies generated by PCD tech compared to a commercially available mouse Cyc hybridoma antibody.
PCD technology utilizes a combination of advantages to offer scientists more effective, reliable, and reproducible means of quickly obtaining high-quality antibodies
PCD technology is also compatible with producing antibodies from other species such as Chickens, or Llamas, each with their own unique and desirable properties.
Figure 1. Rabbit anti-Cyc monoclonal antibody generated by PCD tech. Cyc, a small molecule anti-cancer drug was conjugated to KLH and immunized a rabbit. Immunized rabbit splenocytes were isolated for antibody development with PCD tech. From sixty-one positive clones, four randomly picked clones were tested with EC50 assay. The positive control antibody (blue) is an anti-cyc antibody from a mouse hybridoma line.