In general, conventional mammalian cell libraries are generated by directly inserting DNA of antibodies into a mammalian cell vector. To efficiently screen for fully functional IgG antibodies against specific targets, we sought to integrate the phage and mammalian cell platforms by combining a pre-enriched antibody output and a FACS step for selection. The lower transformation efficiency of mammalian cells has been considered a significant barrier to the construction of highly diverse libraries.Īs described above, both phage and mammalian cell display have inherent limitations that preclude the successful selection of antibodies with desired affinities and low immunogenicity. However, one major limitation of this method is the reduced library diversity it allows compared to phage display. These properties render mammalian cell display extremely attractive and potent for antibody therapeutics development. It enables the display of whole IgG and selection of positive clones with high affinity and other specific biological functions by fluorescence-activated cell sorting (FACS), allowing for highly controlled and real-time selection of IgG. A possibility to address one of these issues is to use mammalian cell display, which possesses intrinsic abilities to fold and glycosylate full-length IgG. Most importantly, the panning process of phage display selection is a black box process, which cannot guarantee the quality of the output clones during the process. This drawback renders a step of conversion to whole IgG molecules and the expression of the candidate clones in mammalian cells necessary for further characterization of their biological activities. Furthermore, antibodies expressed on phages do not undergo normal mammalian posttranslational modifications. One inherent limitation of phage display, however, is that panning a non-immune phage display antibody library sometimes results in the selection of antibodies with K D ranges >100 nM, so it is frequently not sufficient for effective clinical use. It allows for affinity enrichment selections from an extremely large collection of antibodies with sequences of up to 10 11 in size. Phage display is the first and the most widely used platform for the discovery of fully human antibodies. Generating high-affinity antibodies against important drug targets for clinical use remains a challenging task.ĭevelopment of therapeutic antibodies has been accelerated by in vitro antibody selection technologies, which permits rapid generation of millions of clones. Among therapeutic antibodies, antibody affinity plays an important role in biological efficacy, and higher antibody affinity typically allows for lower dosage. The development of mAbs remains a key issue in meeting the world’s profound need for biological drugs. Over the past decades, monoclonal antibodies (mAbs) have become the most important class of therapeutic biologicals on drug market.
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