Rabbit Monoclonal Antibody by Phage display
BIOTEM proposes Rabbit Monoclonal Antibody Development and Production with the Phage Display Technology.
Advantages of Rabbit Monoclonal Antibody:
- High affinity and Specificity
- Recognition of a broad diversity of targets (smaller epitopes, phospho-epitopes, haptens, etc.)
- Somatic mutations in the VH and VL are more frequent
- CDR3-loop of the light chain is longer
Rabbit monoclonal antibodies are therefore of greatest interest for research and diagnostic applications.
BIOTEM does not claim any intellectual properties nor any other rights on the developed antibodies.
- Detection of small molecules (haptens) and non-protein molecules
- Development of immunoassays (low limit of detection)
- Post-translational modification detection (phosphorylation, acetylation, etc.)
- Immunohistochemistry / Immunofluorescence
- Antigen detection in mouse samples
Project
Specifications
1. Project Specifications
For each project, our team explores with the client the challenges of the project and defines together the specifications.
- The target of interest: membrane protein, peptide, small molecule, post-translational modifications, etc.
- Applications: ELISA, WB, FACS, IF, IHC, IP, functional tests, diagnostic application, etc.
- Specifications: Available material, specificity, deadlines, etc.
Taking into account these multiple parameters, BIOTEM proposes tailor-made strategies to meet the client's needs and guarantee the success of the project.
Immunization
Rabbit
2. Immunization
Depending on the client's specifications, BIOTEM has a wide range of proven immunization strategies to obtain high affinity and specificity antibodies. Specific methods are developed to ensure that the appropriate immune response is achieved. At the end of this phase BIOTEM will be able to offer a contract with guaranteed results. The client will also have the possibility to test the sera in his application.
- Protein Immunization : From the production of the recombinant protein (transient transfection in CHO) to its preparation for immunization.
- Peptide Immunization : Immunization with peptides is a relevant strategy for the generation of antibodies directed against selected epitopes. This approach also reduces the need for sophisticated protein preparations for immunization (see also IHC Peptide Profiler®)
- Small Molecule Immunization (Haptens)
- Non-exhaustive list
Library &
Phage Display
3. Library & Phage Display
Unlike naive or synthetic libraries, BIOTEM proposes the construction of an immune library, in scFv format, focused on the target of interest combined with high throughput screening (or bio-panning) by phage display. This optimizes the chances of success in obtaining high affinity and high specificity antibodies.
- Selection of a large number of scFv candidates (VH+VL only)
- Optimized screening with different stringency conditions (direct, competitive, subtractive, etc.)
- Immediate access to sequences (single clones - redundancy analysis)
- Production, Purification and Characterization of candidates (reactivity, affinity, etc.).
At the end of this phase the client will have the opportunity to test the scFv and select the best candidates for the reformatting phase in full Ig format.
Reformatting
Engineering
4. Reformatting
This step allows the transition from scFv to full immunoglobulin format (or orther formats : scFv-Fc, etc.) thanks to a fully integrated platform for the production of recombinant antibodies in mammalian cells (CHO). More info
- Several Isotypes
- Sequence Optimization
- Antibody Engineering (Isotype, mutations, Fc-fusion protein, Bispecific antibodies, Fab, etc.)
- Variants (Chimerization, humanization,etc.)
- Production & Purification (pilot study).
The CLIENT will decide which candidates to produce in large scales.
Production &
Purification
5. Production & purification
Large scale productions are available with our CHO cell Platform. More info
- From milligram to several grams
- Low endotoxin Conditions(< 10 EU/mg ; < 1 EU/mg)
- Serum free System
- Quality Control (concentration, purity, affinity, aggregation, stability, etc.).
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