AMB Express | Japanese scholars optimized and revealed a system platform for screening nanoantibodies using yeast surface display technology
发布时间：
2023-04-14 23:15
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Yeast surface display is a powerful technique for separating and modifying proteins to enhance their activity, specificity, and stability. In this method, gene expression is regulated by promoters, and secretion efficiency is influenced by secretion signals. In addition, the accessibility and activity of displayed proteins are influenced by the length of anchor proteins.
Japanese scholars published an article titled "Evaluation of the yeast surface display system for screening of functional nanoantibodies" in the AMB Express journal, revealing a system platform for screening nanoantibodies using yeast surface display technology.
The ideal combination of promoter, secretion signal, and anchor protein depends on the protein of interest. In this study, we optimized the yeast surface display for nanoscale evaluation. We designed five display systems that use different combinations of promoters, secretion signals, and anchor proteins. Anti hen protein lysozyme nanosomes are used as model nanosomes. The number of nanoantibodies displayed on yeast cells, the number of antigens bound to the displayed nanoantibodies, and the display efficiency were quantified. Overall, we have improved the yeast display system for nanobody engineering and proposed optimization suggestions.
In order to compare the display systems, we attempted to determine the optimal cultivation conditions for them. Both the anti lysozyme nanobody strain and the control strain were cultured at 25 ° C or 30 ° C for 24 hours. At each sampling point, yeast cells were stained and analyzed by flow cytometry (Figure 2a). The control strain showed almost no non-specific absorption of HA labeled antibodies and AF647 labeled lysozyme (Figure 2b). In time history analysis, almost all systems had the highest proportion of yeast cells in the Q2 region within 12 hours (Figure 2c-g). Therefore, five display systems were compared after 12 hours of cultivation.
In this study, we attempted to establish a yeast display system suitable for nanoantibodies. We designed five display systems to study the effects of different promoters, secretion signals, ankyrin and culture temperature. We quantified the number of nanoantibodies displayed on yeast cells, the number of antigens bound to the displayed nanoantibodies, and the display efficiency, and observed that the values of these main endpoints largely depend on the parameters.
