Custom rabbit monoclonal antibody discovery uses single plasma cell interrogation technology and this screens the immune repertoire of rabbits. Rabbits’ monoclonal antibodies are widely recognized and used in research and diagnostics. This is because they are ideal for immunohistochemistry (IHC), flow cytometry, and immunofluorescence (IF). Rabbit monoclonal antibodies can be derived from the traditional method and technological advancement method. Traditionally, rabbit monoclonal is made from rabbit hybridoma which is a process of fusing antibody-secreting cells with a rabbit plasmacytoma cell line. With the technology advancement method, the recombinant rabbit monoclonal antibody can be produced through the use of a phage display library, next-generation sequencing of B cells, and LC-MS/MS of polyclonal antibody.
What are monoclonal antibodies?
First, antibodies are proteins that stick to specific proteins known as antigens. Antibodies revolve around the body system to locate the antigens and attach to them. Once they get attached to it, they force other parts of the immune system to destroy the cells that are containing the antigens. Antibodies can be developed by researchers in the lab to target specific antigens, making them a replica of the natural antibody in the body system. The copies of these antibodies being made in the lab are known as monoclonal antibodies (mAbs or Moabs).
Monoclonal antibodies are man-made proteins acting as human antibodies in the immune system and can bind to specific targets in the body such as antigens. There are different types of monoclonal antibodies and each is developed to bind to a specific antigen. Monoclonal antibodies are developed to be used in the diagnosis and treatment of different diseases including some types of cancer.
Monoclonal antibodies have some distinct peculiarities from polyclonal in that they are produced from a single clonal population of antibody-producing cells. Traditionally, the cells have been hybridomas, which are created through the fusion of antigen-specific plasma B cells with an immortalized myeloma cell. For the production of antibody fragments from non-hybridoma host cells, recombinant technology has also been used. With the cell culture-based methods, monoclonal antibodies can be produced with accurate quality and on a larger scale.
Custom rabbit monoclonal antibody discovery
Rabbit recombinant monoclonal antibodies are a novel source of untapped potential. And this is due to the higher diversity that exists within the antigen-binding regions. Rabbits antibodies that are created from the immunized sources have a higher affinity when compared to mouse antibodies. Antigens that are non-immunogenic in mice, rabbits can respond to it and this makes rabbits a great tool for research reagents use, high sensitivity immuno-detection in the diagnostic of assays, and as sources of therapeutic lead candidates. Although, recombinant antibody protocol for rabbits has remained elusive due to intellectual property and barriers in terms of technology.
A fully optimized solution for custom rabbit monoclonal antibody production can be delivered in high-quality rabbit monoclonal antibodies with the affinity, specity, and functionality that is needed for any difficult targets. And this is done in the following ways
- Through antigen design and its preparation with the use of recombinant protein, peptide, small-molecule, and antigens that are cell-based.
- Using rabbit immunization and serum analysis in optimizing the functional serum assays to ensure that antibodies with the appropriate functions are present in the rabbit before the beginning of library construction.
- Through the library construction, the proprietary cloning method that produces library sizes of 109-1011 capturing higher diversity is compared with other methods to engineer phagemid vectors.
- With selections, the most promising clones are identified through phage panning for accurate affinity, specificity, Fab screening, and sequencing. This selection can be carried out with small molecules, peptides, recombinants, or antigens that are cell-based.
- Through lgG production that deals with cloning and transfection of HEK293T that is followed by purification.
- Then through validation by making use of lgG ELISA or flow cytometry to validate the monoclonal antibody function.
Using ecobody technology for custom rabbit monoclonal antibody production services
Ecobody technology is a rapid screening method for monoclonal antibodies. The technique involved in this method entails that the gene that encodes the monoclonal antibody that is isolated from the single B cells was amplified and then synthesized Fab (fragments of antigens binding) antibody by cell-free protein synthesis. And this is to acquire a target antibody that transport intended specificity. Isolating monoclonal antibodies from B cells has been one of the challenges being faced due to low efficiency and technical difficulties. However, with ecobody technology, the difficulties are being overcome and a highly specific monoclonal antibody for the target antigens is being isolated from B cells that are obtained from a small amount of blood within a short time.
Why rabbit is used for monoclonal antibody discovery service
Rabbits have gained more use for the discovery of monoclonal antibodies due to their immune system which is superior and which can generate higher specificity, sensitivity, affinity, and diversity. Rabbit antibody generated a better response than others in more commonly used species in that
- They have a strong immune response and also increase the immune response to limited epitopes
- They have a unique lgG structure that contains an additional disulfide bond for high stability.
- They have high generic heterogeneity
- They have unique B-cells maturation which includes gene conversion and multiple rounds of somatic hypermutation.
- Their large body size facilitates the large collection of antiserum and immune cells.
- They have long revolutionary distance and their immune systems can respond to human and mouse antigens.