Table 1.

Advantages and disadvantages of technologies used to generate monoclonal antibodies.

Technology	Advantages	Disadvantages
Hybridoma
Original technique	-Preserves the native pairing of variable and constant regions gene combination. -Antibody chimerization and humanization methods and transgenic animals can be used to obtain mAbs for therapeutic use in humans. -Antibodies undergo in vivo affinity maturation.	-Known and available antigen targets are needed. -Low efficiency on cell fusion and hybridoma isolation. -It is required a relatively long period for generating the cell line and the selection of a specific hybridoma. -Hybridoma cell lines may be genetically unstable. -Constant risk of cell culture contamination.
B Cell Targeting	-More efficient cell fusion compared to the original hybridoma technique. -Use only B lymphocyte selected by antigen. -Possibility to simultaneously generate at least 3 specific mAbs against different antigens, using a single mouse.	-Electrostatic field applications might be challenging. -High technical expertise is needed.
Stereospecific targeting	-More efficient cell fusion compared to the original hybridoma technique. -Generation of mAbs that recognize native antigen conformations, instead of linear structures. -DNA immunizations are cheaper than the original hybridoma technique and allow the generation of antibodies against complex or non-conventional antigens.	-Might be more time-consuming and expensive than the previous techniques, particularly if cell lines for immunization, cell fusion, and screening steps are not available. -As it is necessary to perform an electric fusion, it also has the disadvantages of the BCT technique.
Antibody phage display	-Animal host is not required. -The screening of a large number of clones increases the chances of generating good mAbs. -Potential to isolate mAbs against toxic and non-immunogenic antigens. -Possibility to redesign natural CDRs for generating mAbs of improved specificity and affinity. -Display libraries are commercially available.	-The diversity of the phage library depends on the bacterial transformation efficiency. -Antibody formats are limited to scFv and Fab. -Building a phage display library is expensive.
Single B cell	-High efficiency in obtaining specific mAbs, compared to hybridoma technology. -Possibility to isolate mAbs from vaccinated or naturally immunized human subjects. -Isolation of native mAbs with the preservation of natural cognate VH and VL pairing. -No need to culture B cells. -Potential to isolate functional mAbs against conformation determinants that are difficult to emulate in vitro. -It is possible to distinguish B cells at different stages of development and differentiation. -In experimental studies, B cells can be isolated from multiple samples without the need to euthanize animals. -Antibodies undergo in vivo affinity maturation.	-Single-cell sorting devices are expensive. -RT-PCR procedures might be challenging. -Antibodies targeting B cell markers are not available for all species.