An optimized yeast display strategy for efficient scFv antibody selection using ribosomal skipping system and thermo resistant yeast

  • Yanrong Jia
  • Ping Ren
  • Shixin Duan
  • Pei Zeng
  • Debao Xie
  • Fanli ZengEmail author
Original Research Paper



Establish a method to restrict unexpected fragments including stop codons in scFv library and generate a thermo resistant strain for screening of thermal stable scFv sequences.


Here, we have constructed a T2A–Leu2 system for selection of yeast surface display libraries that blocks amplification of “stop codon” plasmids within the library, thereby increasing the quality of the library and efficiency of the selection screen. Also, we generated a temperature-resistant yeast strain, TR1, and validated its combined use with T2A–Leu2 for efficient screening. Thus, we developed a general approach for a fast and efficient screening of scFv libraries using a ribosomal skipping system and thermo-resistant yeast.


The method highlights the utility of the T2A–Leu2-based ribosomal skipping strategy for increasing the quality of the input library for selection, along with an optimized selection protocol based on thermo-resistant yeast cells.


Yeast surface display scFv antibody Ribosomal skipping T2A Antibody screen 


Supporting information

Supplementary Table 1—Nter-SfiI and scFv DNA sequences used in this study.

Supplementary Table 2—Oligos used in this study.

Supplementary Figure 1—Map of pCTcon2-derived plasmid pDJ21.

Supplementary Figure 2—Map of T2A-Leu2 plasmid pDJ22.


This work was supported by a Starting Grant from Hebei Agricultural University (to Fanli Zeng; Grant No. 2018KYYJ01) and by a grant from the National Natural Science Foundation of China (to Fanli Zeng; Grant No. 31801039).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

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Supplementary material 1 (DOCX 16 kb)
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Supplementary material 2 (TIFF 412 kb)
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Supplementary material 3 (TIFF 342 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.College of Life SciencesHebei Agricultural UniversityBaodingChina
  2. 2.Shanghai Institute for Advanced Immunochemical StudiesShanghaiTech UniversityShanghaiChina

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