Fig. 1.
Schematic diagram of monoclonal antibody generation from sampling to isolation of antibodies. (A) The antibody from mature B-cells of a healthy donor is referred for a naïve repertoire. Upon infection or immunization, the mature B-cells is activated to produce plasma B-cells and memory B-cells. This antibody pool is referred to as immune repertoire. (B) The natural antibody repertoire in a healthy individual is diversified via V(D)J recombination, and this represents a naïve antibody library repertoire. Upon infection or immunization, the natural antibody repertoire undergoes somatic hypermutation (SHM), class switch recombination (CSR) and a series of secondary mechanisms to form an immune antibody repertoire. (C) While the recombination of immunoglobulin genes in the variable region are primarily responsible for the diversity of the antibody repertoire, random combination of the heavy (VH) and light (VL) chain during library construction further enlarges the diversity of a library repertoire. (D) The diverse VH/VL gene pairs are then displayed on phages to form an antibody library. The phages are exposed to antigen immobilized on various surfaces for selection, usually subjected to 3–5 rounds of selection process. The final pool of antibodies is analyzed through monoclonal ELISA to isolate out monoclonal antibodies with high specificity.