Major advantages of phage display include the virtual independence of the selection process from target characteristics (e.g., immunogenicity or toxicity of the antigen), which allows application to a wide range of antigens, the possibility of accessing rodent cross reactive antibodies for animal models, the full control over the selection conditions and the technological robustness and relative speed. They differ both in their design regarding framework composition and complementarity-determining region (CDR) diversification, 22 as well as their quality, as defined by library size and correct sequences. 9 - 14 Various naïve, 15, 16 immune, 17 semi-synthetic 18 and synthetic 19 - 21 library formats have been described. On the other hand, the mentioned in vivo methods share the advantage of delivering structurally diverse IgM, IgG and IgA antibodies with naturally paired and correctly folded variable Ig heavy and Ig light (VH/VL) chain frameworks.Īmong the in vitro technologies, the methods of choice for the generation of human mAbs are phage, ribosome and yeast display of recombinant antibody libraries. The hybridoma technology 7 using immunoglobulin (Ig) transgenic mouse systems 8 overcomes this limitation but still suffers from the restriction of the target space due to the need for immunogenic sequences and epitopes. 4 - 6 Target specific human B cell material, however, in some cases may not be readily accessible. Fully human mAbs, for example, can be derived from human B cells by immortalization via viral transformation, 2 through the generation of human hybridomas, 3 or through direct cloning of Ig encoding gene transcripts. 1 A diverse set of in vivo and in vitro technologies are currently available for the generation of these versatile molecules. Our results confirm that quality can be built into an antibody library by prudent selection of unmodified, fully human VH/VL pairs as scaffolds.ĭue to their high specificity and broad target space, monoclonal antibodies (mAbs) represent the most important class of biologics and have great potential for both diagnostic and therapeutic applications. Phage selection was performed against various antigens and unique antibodies with excellent biophysical properties were isolated. Care was taken to minimize the occurrence of potential posttranslational modification sites within the CDRs. The framework regions are fixed and the diversified CDRs were designed based on a systematic analysis of a large set of rearranged human antibody sequences. The selection process included multiple parameters, e.g., assessment of protein expression yield, thermal stability and aggregation propensity in fragment antigen binding (Fab) and IgG1 formats, and relative Fab display rate on phage. The variable Ig heavy and Ig light (VH/VL) chain pairs were selected for biophysical characteristics favorable to manufacturing and development. Ylanthia comprises 36 fixed immunoglobulin (Ig) variable heavy (VH)/variable light (VL) chain pairs, which cover a broad range of canonical complementarity-determining region (CDR) structures. This report describes the design, generation and testing of Ylanthia, a fully synthetic human Fab antibody library with 1.3E+11 clones.
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