It is a great challenge to discover novel chemical reactions suitable for biological analysis in a living system.The development of novel protein thiol blocking agents is a crucial need for exploring protein thiol fun...It is a great challenge to discover novel chemical reactions suitable for biological analysis in a living system.The development of novel protein thiol blocking agents is a crucial need for exploring protein thiol functions in protein refolding,signal transduction,and redox regulation.We are always keen on seeking novel chemical reactions applied to endogenous biological macromolecules or protein thiol sensing,blocking,and labeling.In the present work,we have successfully developed a novel agent to block protein thiol by enhanced electron-withdrawing inductive effects.This sensing and blocking process was detailedly monitored by UV-vis,fluorescent spectra,and SDS-Page gel separation.The spectral studies demonstrated that the agent could react ultrafastly with thiol within seconds atμM level.Furthermore,fluorescent imaging in cells and in vivo was further used for the validation of its ability to sensing and blocking thiol,providing evidence of downregulated protein thiols in Parkinson's disease.The enhanced electronwithdrawing inductive effect strategy in this work may provide a general guideline for designing protein thiol agent.展开更多
Nitrocellose(NC)membranes,as porous paper-like substrates with high protein-binding capabilties,are very popular in the field of point-of-care immunoassays.However,generating robust hydrophobic structures in NC membra...Nitrocellose(NC)membranes,as porous paper-like substrates with high protein-binding capabilties,are very popular in the field of point-of-care immunoassays.However,generating robust hydrophobic structures in NC membranes to fabricate microfluidic paper-based analytical devices(μPADs)remains a great challenge.At present,the main method relies on an expensive wax printer In addition,NC membranes very easy to adhere during the printing process due to electrostatic adsorption.Herein,we developed a facile,fast and low-cost strategy to fabricateμPADs in NC membranes by screen-printing polyurethane acrylate(PUA)as a barrier material for defning flow channels and reaction zones.Moreover,hydrophobic barriers based on UV-curable PUA can resist various surfactant solutions and organic solvents that are generally used in immunoassays and biochemical reactions.To validate the feasibility of this PUA-based NC membrane for immunoassays in point-of-care testing(POCT),we further designed and assembled a rotational paperbased analytical device for implementing a multiplexed enzyme-linked immunosorbent assay(ELISA)in a simple manner.Using the proposed device under the optimal conditions,alpha fetoprotein(AFP)and carcinoembryonic antigen(CEA)could be identified,with limits of detection of 136 pg/mL and 174 pg/mL,respectively,which are below the threshold values of these two cancer biomarkers for clinical diagnosis.We believe that this reliable device provides a promising plaform for the diagnosis of disease based on ELISA or other related bioassays in limited setings or remote regions.展开更多
基金supported by the Natural Science Foundation of China(22376216,21778026,21701074,21976209 and 22204127)the program of the Youth Innovation Promotion Association,CAS(2019217)+3 种基金Taishan Scholar Project Special Funding(TS20190962)the Shenzhen Science and Technology Program(JCYJ20210324142612032)the Guangdong Basic and Applied Basic Research Foundation(2021A1515110906)the Natural Science Basic Research Program of Shaanxi(No.2022JQ-106).
文摘It is a great challenge to discover novel chemical reactions suitable for biological analysis in a living system.The development of novel protein thiol blocking agents is a crucial need for exploring protein thiol functions in protein refolding,signal transduction,and redox regulation.We are always keen on seeking novel chemical reactions applied to endogenous biological macromolecules or protein thiol sensing,blocking,and labeling.In the present work,we have successfully developed a novel agent to block protein thiol by enhanced electron-withdrawing inductive effects.This sensing and blocking process was detailedly monitored by UV-vis,fluorescent spectra,and SDS-Page gel separation.The spectral studies demonstrated that the agent could react ultrafastly with thiol within seconds atμM level.Furthermore,fluorescent imaging in cells and in vivo was further used for the validation of its ability to sensing and blocking thiol,providing evidence of downregulated protein thiols in Parkinson's disease.The enhanced electronwithdrawing inductive effect strategy in this work may provide a general guideline for designing protein thiol agent.
基金suppoted by Key Deployment Project of Centre for Ocean Mega-Research of Science,Chinese Academy of Sciences(COMS2019J01)the National Natural Science Foundation of China(41776110,21876066,22106179)+3 种基金the Shandong Provincial Natural Science Foundation Key Project(ZR2020KB022)the Shandong Provincial Natural Science Foundation Key Research and Development Project(2020CXGC010704)the National Research Foundation of Korea(2019R1A2C3004375,2020R1A5A1018052)the Open Fund of CAS Key Laboratory of Marine Ecology and Environmental Sciences,Institute of Oceanology,Chinese Academy of Sciences(KLMEES202002).
文摘Nitrocellose(NC)membranes,as porous paper-like substrates with high protein-binding capabilties,are very popular in the field of point-of-care immunoassays.However,generating robust hydrophobic structures in NC membranes to fabricate microfluidic paper-based analytical devices(μPADs)remains a great challenge.At present,the main method relies on an expensive wax printer In addition,NC membranes very easy to adhere during the printing process due to electrostatic adsorption.Herein,we developed a facile,fast and low-cost strategy to fabricateμPADs in NC membranes by screen-printing polyurethane acrylate(PUA)as a barrier material for defning flow channels and reaction zones.Moreover,hydrophobic barriers based on UV-curable PUA can resist various surfactant solutions and organic solvents that are generally used in immunoassays and biochemical reactions.To validate the feasibility of this PUA-based NC membrane for immunoassays in point-of-care testing(POCT),we further designed and assembled a rotational paperbased analytical device for implementing a multiplexed enzyme-linked immunosorbent assay(ELISA)in a simple manner.Using the proposed device under the optimal conditions,alpha fetoprotein(AFP)and carcinoembryonic antigen(CEA)could be identified,with limits of detection of 136 pg/mL and 174 pg/mL,respectively,which are below the threshold values of these two cancer biomarkers for clinical diagnosis.We believe that this reliable device provides a promising plaform for the diagnosis of disease based on ELISA or other related bioassays in limited setings or remote regions.
