Discover the highest affinity/specificity Clones

Rabbit monoclonal antibody service (recombinant discovery)

With our patented microfluidics plasma cell discovery (PCD) platform, you can get fully validated, affinity matured rabbit monoclonal antibodies:

  1. Discover top 0.001% high affinity/specificity clones
  2. Pay minimal upfront fees
  3. Deliver in less than 6 months

The scale of phage display meets the power of in vivo affinity maturation--antibody discovery has never been so robust.

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Technology overview

Difficult targets? No problem! Boster Bio has you covered with rabbit mAbs of the highest affinity and specificity tailored for your desired functionality.

Technology Principle

Screen As Early As Possible, Screen As Much As Possible

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.

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Why is Boster’s custom rabbit monoclonal antibodies technology superior?

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.

rabbit-monoclonal-development-service

"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

We promise You a concierge sourcing experience

Scholars serving scholars

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.

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Key concepts/benefits

3 core concepts, move your mouse over to see details.

Recombinant antibody is good

Optimal for Diagnostics and Therapeutics

When building an antibody for diagnostic assays or therapeutic candidate, recombinant antibody is the go to choice. It has the following benefits:

  1. Sequence is available for downstream development such as humanization.
  2. It is stable and scalable.
  3. Its sensitivity and specificity can be further improved by antibody engineering.

Rabbit Is Better Than Mouse

Three Advantages of Rabbit Monoclonals

  1. Higher Affinity and Specificity – Rabbit MAbs typically have 10^3 fold higher affinity between an antigen and antibody than Mouse Monoclonals, ensuring the highest sensitivity and specificity.
  2. Diverse epitope recognition - The lower immune dominance, extensive somatic gene conversion, and a bigger repertoire for hypervariable regions that code the CDRs in the antigen-binding site characterizes the rabbit immune response results in a more diverse epitope recognition yielding a wider range of antibodies.
  3. Improved immune response to small-size epitopes – A unique immune system allows Rabbits to develop antibodies against small moieties in small molecules, lipids, and polymers, or subtle changes in epitopes such as post-translational modifications or single amino acid substitutions.
For more details go to Rabbit Monoclonal page

Finding rare population is key

Screen as much and as early as possible

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.

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Our technology vs. alternatives


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
Cost +++High +++High ++Mid +Very low +++High
Scalability +++High +++High
+++High +Low +++High
Turnaround +Slow +Slow
+Slow +++Fast +Slow
Applications Diagnostics & Therapeutics Diagnostics & Therapeutics
Diagnostics & Research Research Diagnostics & Therapeutics

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.

SERVICE DETAILS

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.

Service overview

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.

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Timeline and costs

Phase 1 Duration Deliverable
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
Phase 2 Duration Deliverable
First Screening with FACS (> a million B cells)
  1. Antigen-specific plasma cell identification and isolation.
  2. Single plasma cell heavy and light chain amplification
2 weeks
Supernatant production 0.25 ml scale, and Secondary screening with ELISA 1 week ELISA data
Phase 3 Duration Deliverable
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
Total 4-5 months

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Deliverables

What you will receive upon successful completion of above phases.

  • ELISA data from both sera and supernatants assays will be delivered.

  • Sequence information of the final delivered clones will be provided, as well as the map of the vector.

  • About 0.5 mg (0.1-1mg) of purified antibody from the two chosen clones to further testing.

  • About 100 ng of heavy and light chain plasmid DNA will be delivered for the final chosen clone.

  • A summary report will be provided

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ADDITIONAL SERVICES

Contact your project concierge for more information.

  • affinity maturation

  • humanization of non-human antibodies

  • epitope mapping

  • scale up production

  • serum free production

  • custom application validation

  • serum free production

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About Boster Bio And PCD

Use for research, IVD and therapeutics

1000+
PCD Projects

Making Antibodies
since 1993

30,000+
publications

Patented
technologies

2000+
RabMabs Available

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Case Study

Antibody Generated with PCD Technology

molecule-doctor-boster

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.

Method

  • Cyc conjugated to KLH was injected into a rabbit to induce an immune response
  • 2 million Rabbit splenocytes were screened for antibody development with PCD technology
  • 94 antigen-specific plasma cells were isolated
  • Sixty-five heavy and light chain genes from these isolated cells were cloned, of which 61 (93.8%) were shown to be antigen-specific by an ELISA.
  • 4 randomly selected rabbit anti-cyclophosphamide monoclonal antibody producing clones, were compared to a commercially available mouse hybridoma normally used in this assay.

Results

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.

plot-od-vs-antibody-concentration

Figure 1. Rabbit anti-Cyc monoclonal antibodies generated by PCD tech compared to a commercially available mouse Cyc hybridoma antibody.

Conclusion

PCD technology utilizes a combination of advantages to offer scientists more effective, reliable, and reproducible means of quickly obtaining high-quality antibodies

  • Isolation of antigen-specific plasma cells
  • Mature antibody selection and single cell amplification
  • Higher diversity and affinity with Rabbit Monoclonals

PCD technology is also compatible with producing antibodies from other species such as Chickens, or Llamas, each with their own unique and desirable properties.

buff-doctor-boster

Other Antibody Tests & Services

FAQs

Q1. What is provided with the Boster custom Rabbit Monoclonal service?

The final deliverable includes ELISA data from both sera and supernatant assays, sequence information and the vector map of the final clones, ~0.5mg purified antibody from two chosen clones for further testing, and approx. 100ng heavy and light chain plasma DNA for the final chosen clone.

Q2. How can you scale up the clones once discovered?

Once the initial clone is discovered, Boster can reproduce anywhere between 1-4mg antibody at any given time, at a starting cost of $300/mg.

Q3. Is the service compatible with other monoclonal hosts?

The PCD technology is entirely compatible with producing antibodies from Chickens, or Llamas, each with their own unique and desirable properties.

Q4. How many antibody clones can be produced using this service?

As many as you would like. We would generally recommend 2 clones

Q5. If I want to raise antibodies to a particular protein, how much protein do I need to send and at what concentration?

A minimum of 1 mg of protein is needed at a concentration of at least 100 µg/mL.

Q6. What guarantee can you give on yield and performance of the Rabbit Monoclonal antibodies developed using the Boster PCD technology?

We will make our best effort to produce the antibody you request. Please note that we cannot guarantee the final yield or that the final product will be biologically active. We do not refund payments for services performed.

Q7. Will a Clone ID be assigned to the clones generated by Boster’s PCD technology?

An in-house antibody clone ID will be assigned to each monoclonal antibody clone that is produced via Boster’s PCD technology, based on its position in the 96 well plate. If the client would prefer to name the clone this option is also available.

Q8. What Is The Recommended Concentration I Should Use For The Primary Antibody? How Do I Decide The Suitable Antibody Dilution?

The end user will need to determine the optimal dilution ratios because of different detection devices, methods, samples, and other factors.

Q9. When Can I Receive The Products After I Place An Order??

The lead time for Custom rabbit monoclonal antibodies usually takes 4-6 months. Please visit our custom rabbit monoclonal antibody service page and contact us for more details about the service and production timeline.

Q10. How Do You Ship Antibodies?

We ship antibodies through FedEx Priority and provide the tracking number to you. The package will include wet ice.

Q11. How Should I Aliquot The Antibodies?

Aliquot volumes are dependent on the amount needed for your experiment. However, we recommend aliquots should 10μL or more. If an aliquot has too little volume, antibody concentration may be affected by liquid evaporation and/or container adsorption.

Q12. How Should I Store The Antibodies? What Is The Shelf Life Of BosterBio's Antibodies?

Our rabbit monoclonal antibodies can be stored at -20°C for one year. After reconstitution, the rabbit monoclonal antibody should be stored at 4°C for one month. If aliquotted, the antibody can be stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.

Q13. How do clones produced by Boster’s custom Rabbit Monoclonal PCD technology compare to commercially available clones?

An example: A Hapten Rabbit Antibody Generated with Boster PCD Technology

Cyclophosphamide (Cyc), a small molecule anti-cancer drug used in cancer chemotherapy, has side effects such as vomiting, hair loss and bleeding from the bladder. Monitoring cyclophosphamide metabolism in the blood can guide doctors to adjust to the proper dosage and minimize side effects. An immunological assay was developed to monitor cyclophosphamide levels in the blood. Cyclophosphamide was conjugated to KLH and injected into a rabbit to induce an immune response. After 3 booster shots, rabbit splenocytes were isolated for antibody development with PCD technology.

Starting with 2 million splenocytes, we isolated 94 antigen-specific plasma cells. Sixty-five heavy chain and light chain genes from these isolated cells were cloned, of which 61 (93.8%) were shown to be antigen-specific by an ELISA. Below, we show the EC-50 curves of 4 randomly selected rabbit anti-cyclophosphamide monoclonal antibody producing clones, compared to a positive control (a mouse hybridoma line commercially available).

rabbit-anti-cyc-monoclonal-antibody

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.


We can safely conclude from this that all four of our 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 in general, and our PCD technology allows us to screen potential candidates with much greater efficiency.

By combining these advantages — Efficient isolation of antigen-specific plasma cells for mature antibody selection, Single cell amplification for an efficient workflow, and utilizing rabbits for more diverse and higher-affinity antibodies — the PCD approach offers scientists a more effective, reliable, and reproducible means of quickly obtaining high-quality antibodies. Another advantage of PCD technology is that it is entirely compatible with producing antibodies from Chickens, or Llamas, each with their own unique and desirable properties.