In Vitro Selection of Single-Domain Antibody (VHH) Using cDNA Display

  • Naoto NemotoEmail author
  • Shigefumi Kumachi
  • Hidenao Arai
Part of the Methods in Molecular Biology book series (MIMB, volume 1827)


Single-domain antibody (e.g., Nanobody, VHH antibody) is a promising scaffold for therapeutic and diagnostic reagents. To expand the range of target molecules, in vitro selection using cell-free display technologies such as cDNA display is useful and powerful because of their huge libraries and robust stability. We provide technical details for in vitro selection of single-domain antibodies using cDNA display.

Key words

cDNA display mRNA display In vitro selection Nanobody VHH Cell-free translation Puromycin Evolutionary molecular engineering Directed evolution Protein engineering Antibody engineering 



The authors thank previous members of the Nemoto Laboratory who helped to develop this protocol, especially Dr. Yuki Mochizuki, Mr. Shinnosuke Kimura, and Mr. Takeru Suzuki. This work was supported by JSPS KAKENHI Grant Number T15H05723 and Brain and Body System Science Institute (BBSSI), Saitama University.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Naoto Nemoto
    • 1
    • 2
    Email author
  • Shigefumi Kumachi
    • 2
  • Hidenao Arai
    • 2
  1. 1.Graduate School of Science and Engineering, Saitama UniversitySaitamaJapan
  2. 2.Epsilon Molecular Engineering, Inc.SaitamaJapan

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