The unique ontogenetic mechanism of rabbit B cells has created a high affinity, high specificity and stable rabbit monoclonal antibody (mAb). The affinity level of rabbit mAb at the picomolar level provides higher sensitivity for clinical diagnosis and treatment, and reduces the dosage of antibody while reducing the side effects of drugs. In addition, the excellent recognition ability of small molecular substances and fine epitopes also makes rabbit monoclonal antibodies more and more popular in the development of immunodiagnostic reagents and antibody drugs, especially in immunohistochemistry, tumor detection and targeted therapy.
DetaiBio provides Rabbit SingleB® mAb Discovery Service, without the commercial authorization of rabbit myeloma cells, in which rabbit antigen-specific memory B cells and plasma cells can be directly screened. It only takes 49 days to obtain more than 50 antibodies that have been verified by FACS assay.
Highly Efficient
whole process lasts for 1.5 months
100 days less than hybridoma
No Commercialization Restrictions
Direct screening of B cells
No hybridoma commercialization restrictions
“1+1” Double Guarantee
PBC&MBC
double screening, double guarantee
High Probability
High antibody diversity
Higher probability of discovery of unique epitopes
| Stage | Service | Timeline | Deliverables |
|---|---|---|---|
| Stage Ⅰ Animal Immunization |
| 48 days | Premium Package
Economic Package
|
| Stage Ⅱ “1+1”Double Screening |
| ||
| Stage Ⅲ BGE® HTP Expression & Functional Test |
| ||
| Stage Ⅳ Monoclonal Antibody Gene Sequencing |
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| Stage Ⅴ Expression and Purification of Recombinant Antibody (optional) |
| For enquiry |
| DetaiBio SingleB® | Homologous Hybridoma | Heterologous Hybridoma | Phage Library | |
|---|---|---|---|---|
| Limitation | High technical threshold | Rabbit hybridoma commercialization restrictions | Poor stability of heterologous fusion | Depending on the correct transcription, translation, folding and assembly of light chains in bacteria |
| Source of Antibody | MBC & PBC | PBC | PBC | MBC & PBC |
| Heavy & Light Chain Pairing | Natural pairing | Natural pairing | Pairing Damage | Unnatural pairing |
| Throughput x Diversity | High | Low | Low | Medium |
| Timeline | 48 days | 5-6 months | 3-4 months | 3-4 months |
| Price | Low | Low | Medium | High |
| Deliverables | Full length antibody sequence & Recombinant plasmid | Hybridoma (easy to degenerate) | Antibody sequence | scFv、Fab、VHH sequence & Recombinant plasmid |
Rapid Immunization of Rabbits
DetaiBio self-developed immunization method with serum titer greater than 1:100K.
Specific MBC Screening
Specific memory B cells were screened by cell surface marker and antigen.
Antibody Functional Verification
The antibody supernatant was obtained by amplification and HTP expression. ELISA was performed on the supernatants and 31 positive clones were found, with a positive rate of 63.3%.
Antibody Affinity Verification
Rabbit monoclonal antibodies discovered by SingleB® Fast mAb discovery platform shows high affinity.
Rabbit IgG has no subclasses, and there are fewer amino acids in the N-terminal and D-E ring, which is easy for molecular cloning and engineering design, and is more suitable for antibody drug development. Rabbit anti 90-95% of light chains are κ 1. There is an in chain disulfide bond between VL and Cl, and the light chain structure is more stable. Rabbit monoclonal antibodies can recognize small molecules, post-translational modifications, single amino acid substitution and other subtle epitope changes, and the specificity of epitope recognition is higher.
Fig1. Schematic diagram of natural rabbit IgG
Note: White part represents light chainκ; Light grey part represents light chain λ; Dark gerypart represents heavy chain; Orange part represents disulfide bond
Rabbits produce antibody libraries through different immune mechanisms from mice and humans, which can produce a strong immune response to foreign antigens, and are also applicable to small molecules and haptens with weak immunogenicity. Human and mice create an anti library through VJ and VDJ gene rearrangement, and then further diversify through somatic hypermutation (SHM). In addition to the SHM mechanism, rabbits can also introduce additional antibody diversity through somatic gene conversion (SGC).
Fig2. Rabbit B cells and antibody library development
Note: GALT: gut-associated lymphoid tissue
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