ProSci is proud to offer: Custom Rabbit Monoclonal Antibody Development & Production
Rabbits are becoming increasingly popular for monoclonal antibody development because of their ability to generate a plethora of diverse antibodies with 10 to 100 times the binding affinity to that of commonly used animal1 antibodies. Rabbit immunization generates a wide variety of immuglobulins, providing the opportunity for identifying distinct antibodies capable of distinguishing between very similar molecules.
Our rabbit monoclonal antibodies are developed directly from isolated plasma cells. The development of these specialized antibodies can help researchers avoid common challenges associated with low affinity and non specific antibodies. ProSci’s rabbit monoclonal antibodies continue to exceed client expectations with wide applicability including diagnostics, therapeutics, and ELISA.
The Difficulties of Rabbit Monoclonal Antibody Development
Current rabbit monoclonal antibody development technologies are limited and time consuming. Traditional fusion of B cells with a myeloma line relies on the B cell secreting the antibody of interest to be fused properly and available for selection during screening. Standard phage display technologies rely on in vitro recombination of heavy and light chains, creating screening and antibodies that are not found in vivo. Traditional direct B cell cloning preserves the original heavy and light chain antibody pairs, but shares a limitation with the other: they use the entire pool of B cells for development, ranging from newly differentiated B cells all the way through fully differentiated B cells, also known as plasma cells. Where as, plasma cells comprise only 0.1-1.0% of the B cell population.
The ProSci Advantage:
Isolated Plasma Cell (IPCTM) Technology
ProSci’s Plasma Cell Technology2 isolates plasma cells at the beginning and uses 100% plasma cells as the starting material for antibody development. This reduces screening time and increases the efficiency of antibodies producing high affinity antibodies. Utilizing our powerful cutting-edge patent in progress technology, ProSci is uniquely positioned to offer customized, specific, high affinity rabbit monoclonal antibodies for your specific needs.
Monoclonal Technology Comparison
|Hybridoma||Phage Display||Single B |
|Single Plasma |
|Start Material||B Cells||B Cells||B Cells||Plasma Cells|
|Effiency||≤ 1% Plasma Cells||≤ 1% Plasma Cells||≤ 1% Plasma Cells||100% Plasma Cells|
|Screening Time||12 Weeks||6 Weeks||3 Weeks||3 Weeks|
Single Plasma Cell Cloning Process
Immunize rabbits on protocol and bleed
• 3 rabbits
Plasma Cell Isolation
Isolate plasma cells
• Isolate plasma cells from 1 rabbit
Gene Amplification & Cloning
Amplify and clone heavy and light chains
• Amplify heavy & light chains
Expression & Screening
Express genes & screen with ELISA
• Plasmid DNA & Sequence for 1 final clone
Optional; Produce large quantities of antibody
• Expression and Protein A purification of additional quantities of antibody
Rabbit Monoclonal Phase Services
Phase I - Immunization: 70-105 Days
- Immunization of 3 rabbits on a 10-week or 15-week protocol using either CFA/IFA or Adjuplex™
- ELISA on each bleed screened against antigen
- Final boost and bone marrow isolation for 1 rabbit
Phase II - Plasma Cell Isolation: 2 Days
- Plasma cell isolation with one cell per well
Phase III - Gene Amplification & Cloning: 7 Days
- Amplification of heavy and light chains from one 96-well plate of plasma cells
- Cloning of the heavy and light chains into expression vectors
Phase IV - Expression & Screening: 11 Days
- Expression of cloned heavy and light chain genes into HEK293 cells
- Screening of supernatants by indirect ELISA against the antigen
- Plasmid DNA & sequence for 1 final clone
- 200 µg of up to 5 clones
Phase V - Antibody Production: 14-28 Days
- Expression and Protein A purification of rabbit monoclonal antibodies from 1 mg to >300 mg
Kurosawa, N., Yoshioka, M., Fujimoto, R., Yamagishi, F., & Isobe, M. (2012).
Rapid production of antigen-specific monoclonal antibodies from a variety of animals. BMC Biology, 10, 80.
Clargo, A. M., Hudson, A. R., Ndlovu, W., Wootton, R. J., Cremin, L. A., O’Dowd, V. L., … Lightwood, D. J. (2014).
The rapid generation of recombinant functional monoclonal antibodies from individual, antigen-specific bone marrow-derived plasma cells isolated using a novel fluorescence-based method. mAbs, 6(1), 143–159.
ANTIBODIES FOR RESEARCH USE ONLY.
A COMMERCIAL LICENSE WILL APPLY FOR COMMERCIALIZATION OF ANTIBODIES FOR THERAPEUTIC USE.