Rabbit Monoclonal Antibodies: Development & Production
Recombinant rabbit monoclonal antibodies are great for targets such as small molecules as they have very high affinity due to the rabbit’s robust immune system and the monospecific nature of monoclonal antibodies. There are many methods to develop custom rabbit monoclonal antibodies such as hybridoma, phage display or B cell cloning. However, unlike other methods, our rabbit monoclonal antibody development process includes directly isolating plasma cells to offer a fully customized, specific, high affinity antibody optimized to your needs. Outsourcing recombinant rabbit monoclonal antibody development to ProSci’s antibody experts allows you to focus on your research.
Isolated Plasma Cell (IPC-TM) Technology
ProSci’s Isolated Plasma Cell TechnologyTM sorts and isolates plasma cells from the beginning, using 100% plasma cells as the starting material for antibody development. With plasma cells as the starting material, the screening efficiency of fully matured antibody clones compared to non-specific clones from immature antibody clones produces high affinity antibodies. Utilizing our powerful cutting-edge technology, ProSci is uniquely positioned to offer custom high affinity rabbit monoclonal antibodies for your specific needs.
Monoclonal Technology Comparison
| Hybridoma | Phage Display | Single B Cell Cloning |
Single Plasma Cell Cloning |
|
|---|---|---|---|---|
| Start Material | B Cells | B Cells | B Cells | Plasma Cells |
| Efficiency | ≤ 1% Plasma Cells | ≤ 1% Plasma Cells | ≤ 1% Plasma Cells | 100% Plasma Cells |
| mAb Diversity | Medium | Low | High | High |
Animal
Rabbits provide strategic benefits over mice when it comes to developing monoclonal antibodies. One rabbit spleen contains more B cells than can be obtained from one mouse spleen. More antibody producing B cells increases the probability of finding the right clone for your needs. Rabbit antibodies also have longer and more heterogeneous CDR3 regions that can improve affinities to targets by 10 to 100 times over murine antibodies. With a unique immune system which reduces immunodominance, they are the ideal host to develop antibodies against a wider range of epitopes, and preferred for developing antibodies against small molecules.
Schedule
Phase
Detailed Procedure
Procedure
I
Immunization
- Immunization of 3 rabbits on 15-week protocol
- ELISA testing of serum
- Final boost and spleen isolation
II
Plasma Cell Isolation and Sorting
- Plasma cell isolation from one rabbit
- Amplification of heavy and light chains
- Cloning into expression vectors
- Express cloned heavy and light chain genes in HEK293 cells
- Screening of supernatants by indirect ELISA
- Supernatants from ELISA-positive wells
III
Deconvolution and Final Expression
- Deconvolute ELISA positive antibody clones
- Purify antibody from 30 ml of culture for up to 4 clones
- Sequencing of two final clones
Antigen Requirements
Antigen Purity
- >90% antigen purity is recommended to ensure effective conjugation, specificity, and higher affinity.
Protein Antigen Specifications
- Quantity 2.5 mg without tag at concentrations of 1 mg/ml or higher required for immunization.
- Size At least 10 kDa
- Production method Use of native protein is preferred over recombinant protein; if a recombinant protein is provided, use an expression system similar to the origin of protein (e.g. use a mammalian expression system for human or other mammalian proteins).
- Tags It is recommended not to have tags for immunization and large fusion proteins such as GST or MBP must be cleaved.
Protein Antigens in Solution
- Concentration Antigens should be submitted at a concentration of 1 mg/ml or higher.
- Buffers Buffers such as PBS, Tris, <4M urea are preferred; SDS and citrate buffers are also acceptable. Please note for services such as ELISA and immuno-affinity purification the antigen needs to be soluble.
- Information required
- buffer
- concentration
- total volume
- total amount
Please submit an antigen submission form with the information above prior to sending your antigen.
Peptide Antigen Specifications
- Quantity
- 5 mg for us to conjugate, or
- 1 mg conjugated to a carrier protein (KLH preferred), and
- 2-3 mg unconjugated for screening
Other Antigen Types
Protein antigens are preferred in rabbit monoclonal, for other antigen types refer to our antigen requirements guide or contact us.
Premier Package
Rabbit monoclonal antibody development using client-provided splenocytes
- Plasma cell isolation and sorting
- Deconvolution and final expression
Pro Package
Rabbit monoclonal antibody development
- 3 rabbit immunization protocol
- Plasma cell isolation and sorting
- Deconvolution and final expression
Add-ons
Additional Small-Scale Purified Antibody If additional purified antibodies are needed, we offer purified antibody from 30 ml culture for additional available clones during the selection process.
Additional Final Clones If you want more final clones than the 2 offered in the packages, we offer additional clones, which include the antibody sequence and DNA.
Differential Screening If you need additional controls during screening, we offer sorting with a second probe (positive or negative), additional ELISAs and ELISA data, to help select the desired clone.
Additional ELISA Screening If you need additional ELISA screening of samples, we offer ELISA screening and data.
Our Assurance
Because of the nature of developing antibodies from immunization to final clones, we implement project milestones consisting of clear, actionable go/no-go decision points during the development process. Our experts review the antibody development progress at each milestone to assure your antibody success.
Custom Rabbit Monoclonal Antibodies
Once antibody development is complete, antibody production can start. We offer 1 L transient expression of rabbit monoclonal antibodies in a mammalian expression system to produce larger quantities of purified rabbit monoclonal antibodies.
Whether your research involves therapeutics, diagnostics, or any other area of study, ProSci offers specialized custom recombinant rabbit monoclonal antibody services for your consideration. If you are looking for more from your current recombinant antibody production service or are looking to make the most out of your research, the choice is clear: ProSci’s custom recombinant antibodies. No other biotechnology firm offers the value of service in customized recombinant antibody development and production.
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