Abstract
Antigen-specific rabbit monoclonal antibodies (RaMoAbs) are useful due to their high specificity and high affinity, and the establishment of a comprehensive and rapid RaMoAb generation system has been highly anticipated. Here, we present a novel system using immunospot array assay on a chip (ISAAC) technology in which we detect and retrieve antigen-specific antibody-secreting cells from the peripheral blood lymphocytes of antigen-immunized rabbits and produce antigen-specific RaMoAbs with 10–12 M affinity within a time period of only 7 days. We have used this system to efficiently generate RaMoAbs that are specific to a phosphorylated signal-transducing molecule. Our system provides a new method for the comprehensive and rapid production of RaMoAbs, which may contribute to laboratory research and clinical applications.
Highlights
Monoclonal antibodies are widely used in laboratory research as well as clinical applications due to their high specificity and high affinity
We immunized a rabbit with hen egg lysozyme (HEL) and prepared IgG+ lymphocytes from peripheral blood lymphocytes (PBLs) (Figure 1A)
We have demonstrated a novel screening procedure for the production of rabbit monoclonal antibodies (RaMoAbs) using the rabbitISAAC system (Figure 1)
Summary
Monoclonal antibodies are widely used in laboratory research as well as clinical applications due to their high specificity and high affinity. Rabbit antibodies generally exhibit a high affinity and high specificity [1,2,3,4,5,6]. Mouse-rabbit hetero-hybridomas were initially used to produce RaMoAbs [10,12,13,14]. These hetero-hybridomas were highly unstable, difficult to clone, and unable to secrete antibodies for prolonged periods [2]. The hybridoma method is not widely used at the laboratory level
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