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A HiBiT/LgBiT-based single-cell platform enables high-throughput screening of extracellular rHuPH20 in Pichia pastoris
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Yifan WuJ
Jie LaiX
Xueting PengL
Lewen HuT
Tao ZhouX
Xinyao Lu* DOI：10.1016/j.bej.2026.110229.png)
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Abstract
En 中文
• A high-throughput single-cell screening platform for secretion-related performance was established in P. pastoris. • N-terminal mutations improved secretion-related performance of rHuPH20. • QTY-based design reduced C-terminal hydrophobicity and improved secretion-related output. • Combined N and C-terminal engineering increased surface display and extracellular enzymatic activity. • Surface fluorescence serves as an effective proxy for secretion performance, enabling enrichment of high-secretion variants.
Keywords:
single-cell screening
Pichia pastoris
rHuPH20
secretion performance
surface fluorescence
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3.8
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6.2K
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