High-throughput extracellular pH monitoring and antibiotics screening by polymeric fluorescent sensor with LCST property

Pan，Tingting1,2; Yang，Cheng2; Li，Jiaze2; Jiang，Jiapei2; Wen，Jiaxing2; Wang，Zijin2; Zhong，Ke2; Tian，Yanqing2; Chen，Meiwan1

2019

10.1016/j.ymeth.2019.04.017

METHODS   影响因子和分区

1046-2023

1095-9130

Monitoring extracellular pH (pHe) is important for biology understanding, since pHe and its homeostasis are closely relevant to cellular metabolism. Hydrogel-based pHe sensors have attracted significant attention and showed wide application, while they are tedious with significant time-cost operation and reproducibility variations for high-throughput application. Herein, we synthesized two polymers for pHe monitoring which are soluble in water at room temperature with easy operations and high reproducibility among various micro-plate wells for high-throughput analysis. P1 (P(OEGMA-co-MEO MA-co-pHS)) and P2 (P(OEGMA-co-pHS)) were synthesized via the Reversible Addition Fragmentation Chain Transfer (RAFT) copolymerization of oligo(ethylene glycol) methacrylate (OEGMA), 2-(2′-methoxyethoxy) ethyl methacrylate (MEO MA) and the pH sensitive fluorescence moiety N-fluoresceinyl methacrylamide (pHS). P1 is soluble in water at room temperature (25 °C) while insoluble at the temperature above 33 °C, indicating its feature of lower critical solution temperature (LCST) at 33 °C. Further P1 showed higher pH sensitivity and photostability than P2 (without LCST property) when used at physiological temperature (37 °C). Thus, P1 was chosen to in-situ monitor the micro-environmental acidification of E. coli, Hela and Ramos cells during their growth, and the metabolism inhibiting activity of a representative antibiotic, ampicillin. Cell concentration-dependent cellular acidification and drug concentration-dependent inhibition of cellular acidification were observed, demonstrating that the LCST polymer (P1) is suitable for real-time cellular acidification monitoring as well as for high-throughput drug screening. This study firstly demonstrated the use of a LCST polymeric sensor for high-throughput screening of antibiotics and investigation of cell metabolism.

Antibiotics activity Extracellular pH monitoring Lower critical solution temperature pH Sensors Polymers

SCI

英语

通讯

Research Fund of the University of Macau[MYRG2016-00130-ICMS-QRCM] ; Research Fund of the University of Macau[MYRG2017-00182-ICMS]

Biochemistry & Molecular Biology

Biochemical Research Methods ; Biochemistry & Molecular Biology

WOS:000500364900008

ACADEMIC PRESS INC ELSEVIER SCIENCE

BIOLOGY & BIOCHEMISTRY

2-s2.0-85065147349

Scopus

1.State Key Laboratory of Quality Research in Chinese MedicineInstitute of Chinese Medical SciencesUniversity of Macau,Taipa,Avenida da Universidade,China
2.Department of Materials Science and EngineeringSouthern University of Science and Technology,Shenzhen,No 1088 Xueyuan Blvd, Xili, Nanshan District,518055,China

Pan，Tingting,Yang，Cheng,Li，Jiaze,et al. High-throughput extracellular pH monitoring and antibiotics screening by polymeric fluorescent sensor with LCST property[J]. METHODS,2019,168:51-61.

Pan，Tingting.,Yang，Cheng.,Li，Jiaze.,Jiang，Jiapei.,Wen，Jiaxing.,...&Chen，Meiwan.(2019).High-throughput extracellular pH monitoring and antibiotics screening by polymeric fluorescent sensor with LCST property.METHODS,168,51-61.

Pan，Tingting,et al."High-throughput extracellular pH monitoring and antibiotics screening by polymeric fluorescent sensor with LCST property".METHODS 168(2019):51-61.