Rabbit Monoclonal Antibody Development
Yurogen offers the development of premium rabbit monoclonal antibodies (mAb), including anti idiotypes, anti post-translational modification sites, anti small molecule compounds, and rare epitopes with greater affinity and specificity. Our SMab™ antibody development platform overcomes issues with traditional methods and utilizes cutting-edge immunological techniques along with cell sorting to screen single B lymphocytes from immunized animals. Over the years, we have successfully supplied more than 10,000 different kinds of rabbit mAbs to our domestic and international clients. These antibodies are widely used in biomedical research and development as well as in clinical practices.
Why Rabbit Monoclonal Antibodies?
Proprietary SMab™ PlatformFirst commercial Single B cell based platform for rabbit mAb discovery
Unique ImmunizationExpertise in DNA, VLP , stable cell line and cell membrane extract immunization
Ex vivo culture of B cellsCultured supernatant for high throughput FACS, WB or reporter cell assay etc. to help quickly narrow down positive candidates
Professional Project DesignTailoring the immunization, sorting and screening strategy for each project
Great Experience in Rabbit mAb DiscoveryDeveloped nearly 10,000 different kind of rabbit mAbs
Reliable Animal SourceUSDA and AAALUC certified domestic animal farm
- Phase I: Immunization
- Phase II: : SMab™ platform screening
- Phase III: Rabbit mAb expression and purification
Phase | Service | Description | Timeline | Phase I | Immunization |
- Standard Package: Immunize two NZW rabbits fo 6-9 weeks with 4 injections and 2 bleed collections
- Bleed titer and measured with ELISA
- (Optional) Additional Assays: ELISA, FACS, WB & IHC etc.
- Client selects one (or more) rabbit for splenocytes isolation
- Phase I report
| 6-9 weeks |
Phase IIa | Single B cell Sorting |
- Antigen specific B cell sorting (by FACS) and culture.
- High throughput ELISA screening for cultured cell supernatant
- Cherry pick ELISA positive clones and freeze cell pallets in RNA shield.
- (Optional) Additional high throughput screening with remaining cells supernatant: ELISA counter screening, FACS, mini-WB & IHC,etc.
| 2 weeks |
Phase IIb | VH/VL amplification & LEM construction |
- VH/VL gene amplification from selected positive B cells
- Linear Expression Module (LEM) construction & transfer into HEK293T cells
- ELISA screen for transfected HEK293T cell supernatant
- (Optional) Additional Assays: Blocking ELISA, affinity ranking, epitope mapping, FACS, WB & IHC etc.
| 2 weeks * |
Phase III | Expression plasmid construction & antibody expression |
- Subclone VH/VL gene into mammalian expression vector (pcDNA3.4) from selected LEM clones
- Small scale expression and protein A/G purification of rabbit monoclonal antibodies
- QC for purified rabbit mAbs: SDS-PAGE and/or Mass-spec, SEC-HPLC (optional)
- Validation of purified rabbit mAbs with ELISA, or (optional) FACS, WB & IHC etc.
Final project report
| 3 weeks * |
* This timeline is based on the construction of 20 LEM clones and 5 final rabbit antibodies.