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- W4383186733 abstract "Classical yeast surface display (YSD) antibody immune libraries are generated by a separate amplification of heavy- and light-chain antibody variable regions (VH and VL, respectively) and subsequent random recombination during the molecular cloning procedure. However, each B cell receptor comprises a unique VH-VL combination, which has been selected and affinity matured in vivo for optimal stability and antigen binding. Thus, the native variable chain pairing is important for the functioning and biophysical properties of the respective antibody. Herein, we present a method for the amplification of cognate VH-VL sequences, compatible with both next-generation sequencing (NGS) and YSD library cloning. We employ a single B cell encapsulation in water-in-oil droplets, followed by a one-pot reverse transcription overlap extension PCR (RT-OE-PCR), resulting in a paired VH-VL repertoire from more than a million B cells in a single day." @default.
- W4383186733 created "2023-07-06" @default.
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- W4383186733 date "2023-01-01" @default.
- W4383186733 modified "2023-10-03" @default.
- W4383186733 title "One-Pot Droplet RT-OE-PCR for the Generation of Natively Paired Antibody Immune Libraries" @default.
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- W4383186733 doi "https://doi.org/10.1007/978-1-0716-3279-6_12" @default.
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