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bioRxiv · 10.1101/2020.06.10.143479

Construction of High Content Nanobody Library in Mammalian Cells by Linear-double-stranded DNA Based Strategies

Zhao, Y.·Su, W.·Li, S.
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Abstract

Here, we developed two novel methods to construct high content nanobody library in mammalian cells. For the first method, we employed a linear-double-stranded DNA based AND-gate (LBAG) strategy. Upstream- and downstream-linear-double-stranded DNAs (ldsDNAs), containing identical overlapping sequences, were co-transfected into cultured cell lines to conduct AND-gate calculation to form intact nanobody expression cassette. For the second method, we generated full-length nanobody expression ldsDNA by in vitro ligation of restrict cut up- and down-stream ldsDNAs. Then the ldsDNAs were directly transfected into mammalian cells to express nanobody library. Both methods generated over a million different nanobody sequences as revealed by high-throughput sequencing (HTS).

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Zhao, Y., Su, W., Li, S.. 2020-06-12. Construction of High Content Nanobody Library in Mammalian Cells by Linear-double-stranded DNA Based Strategies. https://doi.org/10.1101/2020.06.10.143479

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