Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase
Obesity is one of the foremost public health concerns. Human pancreatic lipase (hPL), a crucial digestive enzyme responsible for the digestion of dietary lipids in humans, has been validated as an important therapeutic target for preventing and treating obesity. The serial dilution technique is comm...
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| Format: | Article |
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MDPI AG
2023-02-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/13/2/274 |
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| author | Junqiang Yang Yanyan Deng Min Zhang Shilun Feng Sheng Peng Shijia Yang Peirong Liu Gaozhe Cai Guangbo Ge |
| author_facet | Junqiang Yang Yanyan Deng Min Zhang Shilun Feng Sheng Peng Shijia Yang Peirong Liu Gaozhe Cai Guangbo Ge |
| author_sort | Junqiang Yang |
| collection | DOAJ |
| description | Obesity is one of the foremost public health concerns. Human pancreatic lipase (hPL), a crucial digestive enzyme responsible for the digestion of dietary lipids in humans, has been validated as an important therapeutic target for preventing and treating obesity. The serial dilution technique is commonly used to generate solutions with different concentrations and can be easily modified for drug screening. Conventional serial gradient dilution is often performed with tedious multiple manual pipetting steps, where it is difficult to precisely control fluidic volumes at low microliter levels. Herein, we presented a microfluidic SlipChip that enabled formation and manipulation of serial dilution array in an instrument-free manner. With simple slipping steps, the compound solution could be diluted to seven gradients with the dilution ratio of 1:1 and co-incubated with the enzyme (hPL)-substrate system for screening the anti-hPL potentials. To ensure complete mixing of solution and diluent during continuous dilution, we established a numerical simulation model and conducted an ink mixing experiment to determine the mixing time. Furthermore, we also demonstrated the serial dilution ability of the proposed SlipChip using standard fluorescent dye. As a proof of concept, we tested this microfluidic SlipChip using one marketed anti-obesity drug (Orlistat) and two natural products (1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose (PGG) and sciadopitysin) with anti-hPL potentials. The IC<sub>50</sub> values of these agents were calculated as 11.69 nM, 8.22 nM and 0.80 μM, for Orlistat, PGG and sciadopitysin, respectively, which were consistent with the results obtained by conventional biochemical assay. |
| first_indexed | 2024-03-11T09:04:36Z |
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| id | doaj.art-000d1b6681d1444b916c54e5cdbe6cf5 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-11T09:04:36Z |
| publishDate | 2023-02-01 |
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| series | Biosensors |
| spelling | doaj.art-000d1b6681d1444b916c54e5cdbe6cf52023-11-16T19:26:35ZengMDPI AGBiosensors2079-63742023-02-0113227410.3390/bios13020274Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic LipaseJunqiang Yang0Yanyan Deng1Min Zhang2Shilun Feng3Sheng Peng4Shijia Yang5Peirong Liu6Gaozhe Cai7Guangbo Ge8Department of Anesthesiology, Seventh People’s Hospital of Shanghai University of TCM, Shanghai 200137, ChinaShanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, ChinaShanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, ChinaState Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, ChinaDepartment of Anesthesiology, Longhua Hospital Shanghai University of TCM, Shanghai 200032, ChinaState Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, ChinaDepartment of Anesthesiology, Seventh People’s Hospital of Shanghai University of TCM, Shanghai 200137, ChinaState Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, ChinaShanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, ChinaObesity is one of the foremost public health concerns. Human pancreatic lipase (hPL), a crucial digestive enzyme responsible for the digestion of dietary lipids in humans, has been validated as an important therapeutic target for preventing and treating obesity. The serial dilution technique is commonly used to generate solutions with different concentrations and can be easily modified for drug screening. Conventional serial gradient dilution is often performed with tedious multiple manual pipetting steps, where it is difficult to precisely control fluidic volumes at low microliter levels. Herein, we presented a microfluidic SlipChip that enabled formation and manipulation of serial dilution array in an instrument-free manner. With simple slipping steps, the compound solution could be diluted to seven gradients with the dilution ratio of 1:1 and co-incubated with the enzyme (hPL)-substrate system for screening the anti-hPL potentials. To ensure complete mixing of solution and diluent during continuous dilution, we established a numerical simulation model and conducted an ink mixing experiment to determine the mixing time. Furthermore, we also demonstrated the serial dilution ability of the proposed SlipChip using standard fluorescent dye. As a proof of concept, we tested this microfluidic SlipChip using one marketed anti-obesity drug (Orlistat) and two natural products (1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose (PGG) and sciadopitysin) with anti-hPL potentials. The IC<sub>50</sub> values of these agents were calculated as 11.69 nM, 8.22 nM and 0.80 μM, for Orlistat, PGG and sciadopitysin, respectively, which were consistent with the results obtained by conventional biochemical assay.https://www.mdpi.com/2079-6374/13/2/274microfluidicSlipChipserial gradient dilutionobesityhuman pancreatic lipaseinhibitors |
| spellingShingle | Junqiang Yang Yanyan Deng Min Zhang Shilun Feng Sheng Peng Shijia Yang Peirong Liu Gaozhe Cai Guangbo Ge Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase Biosensors microfluidic SlipChip serial gradient dilution obesity human pancreatic lipase inhibitors |
| title | Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase |
| title_full | Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase |
| title_fullStr | Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase |
| title_full_unstemmed | Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase |
| title_short | Construction and Manipulation of Serial Gradient Dilution Array on a Microfluidic Slipchip for Screening and Characterizing Inhibitors against Human Pancreatic Lipase |
| title_sort | construction and manipulation of serial gradient dilution array on a microfluidic slipchip for screening and characterizing inhibitors against human pancreatic lipase |
| topic | microfluidic SlipChip serial gradient dilution obesity human pancreatic lipase inhibitors |
| url | https://www.mdpi.com/2079-6374/13/2/274 |
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