title: Imaging trypsin activity through changes in the orientation of liquid crystals coupled to the interactions between a polyelectrolyte and a phospholipid layer.
abstract: In this study, we developed a new type of liquid crystal (LC)-based sensor for the real-time and label-free monitoring of enzymatic activity through changes in the orientation of LCs coupled to the interactions between polyelectrolyte and phospholipid. The LCs changed from dark to bright after an aqueous solution of poly-l-lysine (PLL) was transferred onto a self-assembled monolayer of the phospholipid, dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), at the aqueous/LC interface. Interactions between the positively charged PLL and the negatively charged DOPG drove the reorganization of the phospholipid membrane, which induced an orientational transition in the LCs from a homeotropic to planar state. Since the serine endopeptidase trypsin can enzymatically catalyze the hydrolysis of PLL, the dark-to-bright shift in the optical response was not observed after transferring a mixed solution of PLL and trypsin onto the DOPG-decorated LC interface, indicating that no orientational transitions in the LCs occurred. However, the optical response from dark to bright was observed when the mixture in the optical cell was replaced by an aqueous solution of PLL. Control experiments with trypsin or an aqueous mixture of PLL and deactivated trypsin further confirmed the feasibility of this approach. The detection limit of trypsin was determined to be ~1 μg/mL. This approach holds great promise for use in the development of LC-based sensors for the detection of enzymatic reactions in cases where the biological polyelectrolyte substrates of enzymes could disrupt the organization of the membrane and induce orientational transitions of LCs at the aqueous/LC interface.
authors.affiliation: Department of BioNano Technology, Gachon University, San 65, Bokjeong-Dong, Sujeong-Gu, Seongnam-City, Gyeonggi-Do, 461-701, Korea.
authors.forename: Qiong-Zheng
authors.initials: QZ
authors.lastname: Hu
authors.forename: Chang-Hyun
authors.initials: CH
authors.lastname: Jang
mesh_terms: D004573:Electrolytes / Q000737:chemistry*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D057230:Limit of Detection; D008051:Lipid Bilayers / Q000378:metabolism*; D050866:Liquid Crystals / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry; D010743:Phospholipids / Q000378:metabolism*; D011107:Polylysine / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism*
chemical_list: D004573:Electrolytes; D008051:Lipid Bilayers; D010715:Phosphatidylglycerols; D010743:Phospholipids; D011107:Polylysine; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol; D014357:Trypsin