Rabbit mAbs Development

Custom Antibody Development

Our team has decades of experience on high quality monoclonal antibody development services. DIMA’s monoclonal antibody (mAb) development platform is a revolutionary technology platform for mAb development. Different from hybridoma fusion platform, we can directly isolate IgG genes from B cells of immunized animals. At present, we have successfully completed hundreds of custom development projects with high success rate. Many of them are raised against the difficult membrane proteins with high affinities and manufacturability for the downstream antibody drug development. 

Service Consultation


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    Service details

    Custom rabbit mAbs development procedure & deliverables

    StepService detailsDeliverablesEstimated production cycle (week)
    Immunogen preparationPlasmid construction, mammalian cell produced immunogens or supplied by customer7-10 Weeks
    Animal immunizationImmunize two rabbits against the same antigenCrude sera and ELISA data on anti-serum titer test5-8 Weeks
    B cell isolation and amplificationPBMC collection and cryopreservation; target positive B cell enrichment and single B cell expansion100ul B cell supernatant;B cell supernatant ELISA test data2-3 Weeks
    DIMA mAbs cDNA cloning, sequencing and storageIgG gene cloning, Expression vector construction; small-scale antibody production and validation; select 1-2 positive clones for full-length IgG sequencing and sequence analysisSequencing report of one clone; ELISA data3-4 Weeks
    DIMA mAbs production and validation Antibody production, purification and QC validation1 mg purified recombinant antibody, antibody QC data3-4 Weeks

    Rabbit Monoclonal Antibody

    Comparing with other animals, the rabbit has a unique B cell development process and is more distant from human than rodent. Besides these, rabbit B cells utilize a dual affinity maturation mechanism, including gene conversion and SHM (somatic hyper-mutation). At the same time, the rabbit IgG has a unique protein structure different from the mouse IgG, including one subtype of IgG, more noncanonical C-C for light chain, great variation in length and sequence for CDR3, etc. These differences might make rabbit produce antibodies with high affinity and wide diversity. In recent years, rabbit monoclonal antibodies are getting popular in different immunological assay development, especially for immunohistochemistry application. So far, 5 FDA approved companion diagnostic IHC antibodies are rabbit monoclonal antibodies (Download). In the past, rabbit monoclonal development technology required a patent protected rabbit myeloma cell line, which limited its applications in many areas. Right now, DIMA’s new proprietary DIMA mAbs technology platform provides a great technology breakthrough in this area. Without using myeloma cells, we can directly isolate IgG genes from positive B cell clones. This will not only enable us to shorten the development time, but also directly acquire the genetic coding information of IgG without worrying of losing hybridoma cells.

    Comparison of rabbit monoclonal antibody and mouse monoclonal antibody

    Comparison ItemsRabbit Monoclonal AntibodyMouse Monoclonal Antibody
    B cell antibody spectrumV (D) J recombination; continued development in Appendix or other gut associated lymphoid tissues, continued to increase the diversity of antibody spectrum and the size of antibody library through very slow gene conversionV (D) J arrangement
    The mechanism of antibody maturationGene conversion + hypermutationHypermutation
    CD1 FamilyFive CD1 isoforms have strong immune responses to antigens such as lipids and glycolipids; recognize polypeptides and small moleculesOne CD1d; many small molecular compounds and polypeptides do not cause a good immune response in mice
    IgG TypeOne IgG with more disulfide bonds, more complex CDR region and light chain structureIgG1, IgG2a, IgG2b, IgG2c, IgG3
    Species responseStronger response to human and rodent antigensWeak response to rodent antigens
    Epitope recognitionAntibodies can recognize multiple epitopes of antigens and have advantages in post-translational modification (such as phosphorylation, methylation, acetylation, ubiquitination like)Limited epitope recognition by antibody
    AffinityThe Kd value (equilibrium dissociation constant) can usually reach the picomore level (Kd = 10-12M)Kd value is in the range of nanomolar (Kd = 10-9M)
    ProductionThe recombinant expressed antibody is stable between batches, and the automatic preparation process can be establishedThe antibody secreted by hybridoma is unstable, which is easy to cause the loss of antibody gene
    StoreThe antibody gene sequence is obtained directly, and the antibody information is easy to store and transmitThe cost of hybridoma cell preservation is very high, which requires a lot of liquid nitrogen and cell storage equipment
    ApplicationWB, ELISA, FC, IP, IHC, ICC (obvious advantages in IHC)WB, ELISA, FC, IP have no obvious advantages in IHC and ICC

    Applications

    Application in clinical diagnosis
    Recent evidence indicated that rabbit monoclonal antibody has clear advantages in immunohistochemistry application. Currently rabbit monoclonal antibodies become the favorite choices for pathologists. At present, FDA has approved 11 rabbit monoclonal antibodies for diagnosis, including companion diagnostic IHC antibodies for anti-PD-1 and anti-PD-L1 immunotherapy drug treatment, 4B5 clone for Herceptin treatment, etc.

    Part of the FDA approved list of rabbit monoclonal antibodies for companion diagnosis

    Concomitant diagnostic reagentsAntibody drugCancerDetection platform
    PATHWAY ANTI-HER-2/NEU (4B5)HERCEPTIN(trastuzumab)Breast cancerIHC
    VENTANA ALK (D5F3) CDx AssayXALKORI(crizotinib)Non-small cell lung cancerIHC
    PD-L1 IHC 28-8OPDIVO(nivolumab)Melanoma Non-small cell lung cancerIHC
    VENTANA PD-L1 (SP142) AssayTECENTRIQ(atezolizumab)Bladder cancer Non-small cell lung cancerIHC
    VENTANA PD-L1 (SP263) AssayKEYTRUDA®(pembrolizumab)mNSCLCIHC

    Application on therapeutic antibody drug development
    On October 8th, 2019, the first rabbit monoclonal antibody derived drug Beovu® was approved by FDA for the treatment of wet age-related macular degeneration (AMD). Beovu® is a variable region fragment of humanized rabbit monoclonal antibody scFv targeting VEGF. This is also the 83rd antibody drug approved by FDA. Compared with other antibodies, rabbit recombinant monoclonal antibodies have higher affinity, which can not only reduce the clinical dosage of antibodies, but also reduce the side effects caused by the use of a large dosage.

    Application in scientific research
    With the advantages of high specificity and high affinity, rabbit monoclonal antibodies have been widely used in life science research. At present, rabbit monoclonal antibodies have been widely used as reagents for immunoblotting, immunohistochemistry, flow cytometry and other immunology research. Due to its unique B cell development system, rabbit monoclonal antibodies have exhibited special strength in detecting subtle difference on small epitopes, such as post-translational modified amino acids within a protein.

    Service Case

    The whole DIMA mAbs development procedure took around 3.5 months
    (delivered 8 flow working rabbit mAbs to customer (Bioraid, a CAR-T company))

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