In our experience, the level of antibody diversity that can be obtained depends on the organ used for library construction. Specifically, spleen, bone marrow, PBMCs (peripheral blood mononuclear cells) and lymph nodes, and GALT (gut-associated lymphoid tissue) are sources of antibodies. We can obtain a much greater variety of high-performance antibodies using spleen and bone marrow than using PBMC or GALT.
If cost is the limiting factor, we recommend bone marrow as the source of the antibody library. However, if you are aiming to develop antibodies against difficult targets such as GPCRs or specific epitopes with desired functions, we strongly recommend that you generate antibodies from both spleen and bone marrow.
In general, library construction is performed by the PCR amplification of antibody genes utilizing 2 gene-specific primers. This causes biased amplification and non-specific amplification resulting in poor coverage of antibody repertoire in the immunized animals.
AbWiz Bio invented a unique method called WizAmpTM (US 9,890,414) that engineers cDNA specifically for antibody genes and utilizes only 1 non-gene-specific primer (single primer amplification). This method ensures the widest coverage of antibody repertoire including the rare clones that have the desired function and specificity.