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Our motivation for developing zebrafish specific rabbit monoclonal antibodies

Although zebrafish has been a very powerful model in biomedical research, less than 3% of zebrafish proteins have available antibodies, not to mention monoclonal antibodies. Yurogen Biosystems LLC would like to bring our proprietary single cell based SMabTM platform and develop rabbit monoclonal antibodies (RmAbs) to zebrafish proteins to remove this reagent hurdle that hinders many research projects in fast-growing zebrafish field. With the help from zebrafish community, we will rigorously validate our recombinant RmAbs to ensure they will work reproducibly for specific applications such as western blotting or whole mount immunofluorescence staining. In addition, we can deliver the gene sequences of RmAbs to enhance reproducibility and ease antibody engineering for special applications such as generating transgenic zebrafish with specific RmAbs to knock down protein function in a controllable manner. We envision that RmAbs will be important and powerful addition to expand the current zebrafish research resources.

Advantages of rabbit monoclonal antibodies

Unlike mice and other rodents, rabbit has a better immune system which can generate antibodies with greater diversity, higher affinity, broader epitope recognition, and better stability [1]. Therefore, RmAbs have many advantages which set them apart from traditional mouse antibodies. Taking advantage of our robust SMabTM platform, we can quickly and efficiently generate high quality rabbit monoclonal antibodies for a variety of antigens.

High Affinity Typically, RmAbs have 10 – 100 times higher affinity than mouse monoclonal antibodies. On average, the KD values of RmAbs are range from 10-12 M up to 10-14 M [3].

Superior Specificity The development of primary and secondary antibody repertoires in rabbits possesses unique characteristics, such as gut-associated lymphoid tissues (GALT) B cell development and multiple selection procedures. These unique characteristics may account for the improved quality of the antibodies produced in rabbit [4,5]. The high specificity of RmAbs has clear edges in assays: improved assay sensitivity, improve accuracy with decreased false positive rate, and lower background noises. These features make RmAbs ideal choices for stringent assays including IHC in clinics [6].

Broad Epitope Recognition including Haptens Due to its reduced immunodominance, rabbit antigen presenting cells can present broader range of epitopes than their mouse counterparts. Many less abundant proteins or epitopes can be well presented and recognized by rabbits. Rabbits are therefore able to generate high affinity antibodies against difficult epitopes, such as protein modifications, enzyme cleavage sites, or conformational epitopes. Partially due to the expression of all three subclasses of CD1 proteins, the non-peptidic epitope presenting in rabbits is more efficient. As a result, rabbit can handle haptens better than mice and is widely adopted as preferable host to generate better antibodies to haptens, including small molecule drugs, steroid hormones, lipids, and glycolipids.

Great Stability and Long Shelf-life Rabbit IgG has extra disulfide bonds in the variable region of the heavy chain and an extra disulfide bond between Vκ and Cκ. These extra disulfide bonds may result in the great stability and long shelf-life of rabbit antibodies.

 

About Yurogen

Yurogen Biosystems LLC is found in 2015, located in the Greater Boston area. Yurogen strives to become a leading provider of premium monoclonal antibodies and antibody-based products for our customers in both research community and industries. Yurogen has developed an innovative single cell based SMabTM platform that can efficiently screen monoclonal antibodies from a variety of host animal in a cost-efficient manner without sacrificing antibody quality. Since 2015, Yurogen has quickly established and is actively marketing our own product lines of RmAbs. In addition to our own product lines, we offer full scope of antibody CRO services including antigen design and production, and antibody screening, engineering, and production.

 

References:

1. Schiaffella E, Sehgal D, Anderson A, Mage R. Gene conversion and hypermutation during diversification of VH sequences in developing splenic germinal centers of immunized rabbits. J Immunol. 1999;162:3984-95.

2. Pan R, Chen Y, Vaine M, Hu G, Wang S, Lu S, Kong XP. Structural analysis of a novel rabbit monoclonal antibody R53 targeting an epitope in HIV-1 gp120 C4 region critical for receptor and co-receptor binding. Emerg Microbes Infect. 2015 Jul;4(7)

3. Gendusa R, Scalia C, Buscone S, Cattoretti G. Elution of High-affinity (>10-9 KD) Antibodies from Tissue Sections: Clues to the Molecular Mechanism and Use in Sequential Immunostaining. J Histochem Cytochem. 2014;62:519-531

4. Rossi S, Laurino L, Furlanetto A, Chinellato S, Orvieto E, Canal F, et al. Rabbit monoclonal antibodies: a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies. Am J Clin Pathol. 2005;124:295-302

5. Rocha R, Nunes C, Rocha G, Oliveira F, Sanches F, Gobbi H. Rabbit monoclonal antibodies show higher sensitivity than mouse monoclonals for estrogen and progesterone receptor evaluation in breast cancer by immunohistochemistry. Pathol Res Pract. 2008;204:655-62

6. Vilches-Moure J, Ramos-Vara J. Comparison of rabbit monoclonal and mouse monoclonal antibodies in immunohistochemistry in canine tissues. J Vet Diagn Invest. 2005;17:346-50

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