Rabbit monoclonal antibody production
- High affinity & specificity
- Optimal for small molecules and non-protein detection
- Novel epitope recognition
Rabbit Monoclonal Antibody Platform
- Detection of small molecules (haptens) and non-protein molecules
- Development of immunoassays requiring a low limit of detection
- Post-translational modification detection (phosphorylation, acetylation, etc.)
- Immunohistochemistry / Immunofluorescence (more accurate results)
- Antigen detection in mouse samples
Higher SpecificityRabbits generate a highly diverse B-lymphocyte repertoire, especially through their CDR3-loop which is long when compared to other animals.
The combined benefits of a strong natural immune response and a high throughput Phage Display screening allow the selection of antibodies with remarkable specificity. They are optimal for the detection of:
- Small molecules (toxins, pollutants, hormones, drugs, etc.)
- Non-protein targets (lipids, carbohydrates, etc.)
- Post translational modifications (phosphorylation, etc.)
- Point mutations
Higher AffinityThanks to a wider antibody repertoire and a more efficient affinity maturation system, rabbit antibodies frequently exhibit 10- to 100- fold higher affinity values than mice and other rodent monoclonal antibodies.
Affinities values (KD) are often in the nanomolar (10-9 M) range and may go down to picomolar (10-12 M) range without any in vitro affinity maturation.
In addition, higher antibody affinities ensure higher signal-to-noise ratio when compared to mouse mAbs at a given antibody concentration.
Larger Epitope CoverageAntibodies developed in rabbits have generally larger epitope coverage compared to antibodies derived from mice.
Thanks to the long phylogenetic distance of rabbits (Lagomorpha) from humans and mice / rodents, human targets are often more immunogenic in rabbits than in mice.
Moreover the rabbit immune system presents different advantages:
- Lower immune dominance, immune response is directed against various epitopes
- Larger B-cell repertoire
Rabbit Immunization & Library Construction
BIOTEM proposes optimized strategies for immunogen / antigen development as well as custom rabbit immunization protocols.
Specific pathogen-free (SPF) animals are immunized and extensive serum analysis (ELISA, affinity analysis, etc.) is performed. Tailored methodologies are developed to assess that appropriate immune responses have been achieved.
Following an in-depth analysis of the immune responses, BIOTEM selects the best animals in order to build highly representative hyper-immune libraries focused on the antigen.
The combination of a strong natural immune response and an antigen based library construction allows the generation of high affinity antibodies with a large epitope coverage
Focus on « Hapten targets »
Small chemical compounds (haptens) are usually not immunogenic and therefore need to be conjugated to a carrier protein to elicit a strong immune response. Hapten conjugation strategy (hapten orientation, linker characteristics and hapten:carrier ratio) will greatly influence the quality of developed antibodies.
BIOTEM disposes of a chemical unit dedicated to the custom development of immunogens:
- Hapten synthesis
- Hapten functionalization
- Hapten conjugation to carrier proteins
Our expertise allows the development of optimized immunogens for the generation of antibodies with high affinity and specificity.
Phage Display & scFv
Based on these elements BIOTEM implements the best biopanning methods, adapted to the CLIENT’s antigens:
BIOTEM customized biopanning strategies allow the selection of a greater number of binders compared to classical hybridoma technology.
Post-panning isolated binders are individually selected, sequenced and tested for their reactivity / specificity by different methods (ELISA, affinity analysis, …).
- ScFv- or full immunoglobulin- format (various isotypes available)
- Small and large scale batch production
- Custom / adapted purification protocols
- Low endotoxin preparation
- BIOTEM Quality Control
- A comprehensive set of quality controls are available on lead antibodies:
- Concentration determination
- Purity analysis
- Affinity constant determination
- Endotoxin analysis
- Aggregation and oligomerization (AUC, DLS & SEC-HPLC)
- Stability (DSC & Circular Dichroism)
- Other quality controls upon request