rabbit monoclonal Antibody Production Service

Fig. 1 Schematic drawing of natural rabbit antibodies in IgG format. Reference source: Weber, J., Peng, H. & Rader, C. From rabbit antibody repertoires to rabbit monoclonal antibodies.

Compared with mouse derived antibodies, rabbit monoclonal antibodies can recognize more human antigens and can be used for antibody screening against antigens with high homology between humans and mice. The selected rabbit anti mouse antigen antibodies can be used as reference antibodies, and the current application is in the study of mouse stem cell antigens. The most commonly used experimental mice in experiments are usually inbred mice. Therefore, the affinity of mouse monoclonal antibodies prepared by conventional immunization methods is low, but this problem does not occur when rabbits are immunized and monoclonal antibodies are prepared. Due to the unique conformation of CDR3 in rabbit monoclonal antibodies, their affinity may be higher. There are many types of CD1 family proteins in rabbits, and CD1 family proteins are mainly responsible for the transmission of lipids and glycolipids. Therefore, rabbit derived antibodies have a higher affinity for lipids and carbohydrates, and high-quality rabbit antibody production targeting lipids and glycolipids antigens can be achieved. Antibodies are usually screened by isolating B cells from peripheral blood. Rabbits have a larger volume than mice and can provide a relatively higher amount of blood, making them more advantageous for long-term affinity maturation antibody screening. In addition, based on the high affinity and unique structure of rabbit monoclonal antibodies, the probability of successful humanized modification of rabbit monoclonal antibodies is high.

The process of rabbit antibody production first requires selecting immunogens according to demand, mixing the processed immunogens with adjuvants, and then injecting them into rabbits to stimulate the occurrence of immune responses. Rabbits are immunized multiple times to obtain sufficient immune response, with a certain interval between each immune response, usually two weeks, and the dose of immunogen gradually increases. After a period of immunization, B cells were collected from the lymphoid tissue of rabbits, cells that do not secrete antibodies were extracted from myeloma cell lines. The collected B cells were fused with myeloma cells to form hybridoma cells using techniques such as polyethylene glycol (PEG) or electrofusion. The successfully fused hybridoma cells are screened to secrete antibodies against specific antigens. A single clone was selected from the hybridoma cell population and cultured to ensure a single source of antibodies. The selected monoclonal hybridoma cells were cultured in vitro to produce a large amount of antibodies. In addition, antibody amplification can also be achieved by injecting hybridoma cells into host animals such as mice, collecting samples of host animal blood or ascites, and extracting components containing rabbit monoclonal antibodies.

Fig. 2 Screening process of monoclonal antibodies based on B cell culture