● Introduction
The production of antibodies by plasma cells (also known as effector B cells) has first described by Astrid Fagraeus in 1948. And the hybridoma technology, described by Georges Köhler and Cesar Milstein in 1975, is based on the immunization of animals with the desired antigen, followed by the fusion of specific B lymphocytes with “immortal” myeloma cells. Although hybridoma antibody production is a well-established method of producing antibody, there are several drawbacks. They have limited sensitivity and are susceptible to antigen heterogeneity and experimental condition changes. Hybridomas are also known to undergo genetic drift leading to batch-to-batch variability, or they may even lose antibody genes completely arresting the production of antibodies.
RecRAbs® recombinant rabbit monoclonal antibodies manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. This unique technology employs innovative cell sorting, enriching, advanced cloning. It does not take the traditional approach of using the hybridoma method, hence avoiding the stability issue associated with hybridomas.
The workflow includes (i) Inoculation of rabbit with specific antigen (peptide or protein), (ii) identification and isolation of single B cells from rabbit blood expressing IgG antibodies, (iii) an elaborate short term B-cell cultivation to produce sufficient monoclonal antigen specific IgG for comprehensive phenotype screens, (iv) isolation of VH and VL coding regions via PCR from B-cell clones producing antigen specific and functional antibodies followed by the sequence determination, (v) recombinant expression in a host (bacteria, yeast, insect, or mammalian cell lines) and purification of IgG antibodies, (vi) application screening – ELISA, WB, IP, IHC, IF, FCM, etc.
● RecRAbs®——Zenbioscience Rabbit Monoclonal Antibody Production Process
● Advantages of Recombinant Rabbit Mnoclonal Atibody
√ High affinity and superior specificity
√ Broad epitope recognition
√ Excellent lot-to-lot consistency due to the recombinant technology
√ Good recognition of small molecules and post translational modification sites
√ Multiple applications, including IF, IHC, ChIP, ELISA, Flow, IP and WB
√ Animal origin-free formulations, the result of production by cell culture
Zen-bioscience provides a variety of solid validated recombinant rabbit mAbs that can be used in various biochemical experiments such as WB, IHC-P, ICC/IF, etc.