An optimized yeast display strategy for efficient scFv antibody selection using ribosomal skipping system and thermo resistant yeast
| Autor: | Debao Xie, Shixin Duan, Fanli Zeng, Ping Ren, Pei Zeng, Yanrong Jia |
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| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine Ribosomal skipping Hot Temperature Computer science Bioengineering Computational biology Yeast display 01 natural sciences Applied Microbiology and Biotechnology 03 medical and health sciences Plasmid 010608 biotechnology Resistant strain Yeasts Genetic Testing Selection (genetic algorithm) Gene Library Protein Stability General Medicine Yeast strain Stop codon Yeast 030104 developmental biology Metabolic Engineering Cell Surface Display Techniques Biotechnology Single-Chain Antibodies |
| Zdroj: | Biotechnology letters. 41(8-9) |
| ISSN: | 1573-6776 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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