An　all-in-one　paper-based　analytical　device　(PAD)　was　successfully　developed　for　visual　fluorescence　detection　of　carcinoembryonic　antigen　(CEA)　on　CdTe/CdSe　quantum　dot　(QD)-enzyme-impregnated　paper　by　coupling　with　a　bioresponsive　controlled-release　system　from　DNA-gated　mesoporous　silica　nanocontainers　(MSNs).　The　assay　was　carried　out　in　a　centrifuge　tube　by　using　glucose-loaded　MSNs　with　a　CEA　aptamer　and　a　QD-enzyme-paper　attached　on　the　lid.　Initially,　single-strand　complementary　DNA　to　a　CEA　aptamer　was　covalently　conjugated　to　the　aminated　MSN,　and　then　glucose　(enzyme　substrate)　molecules　were　gated　into　the　pore　with　the　help　of　the　aptamer.　Glucose　oxidase　(GOD)　and　CdTe/CdSe　QDs　were　coimmobilized　on　paper　for　the　visual　fluorescence　signal　output.　Upon　target　CEA　introduction　in　the　detection　cell,　the　analyte　specifically　reacted　with　the　immobilized　aptamer　on　the　MSN　to　open　the　pore,　thereby　resulting　in　the　glucose　release.　The　released　glucose　was　oxidized　by　the　immobilized　GOD　on　paper　to　produce　gluconic　acid　and　hydrogen　peroxide,　and　the　latter　quenched　the　fluorescence　of　CdTe/CdSe　QDs,　which　could　be　determined　by　the　naked　eye　on　a　portable　smartphone　and　a　commercial　fluorospectrometer.　Under　optimal　conditions,　the　PAD-based　sensing　system　enabled　sensitive　discrimination　of　target　CEA　against　other　biomarkers　or　proteins　in　a　linear　range　of　0.05-20　ng　mL-1　with　a　limit　of　detection　of　6.7　pg　mL-1　(ppt).　In　addition,　our　strategy　displayed　high　specificity,　good　reproducibility,　and　acceptable　accuracy　for　analyzing　human　serum　specimens　with　a　commercial　human　CEA　ELISA　kit.　Importantly,　this　methodology　offers　promise　for　simple　analysis　of　biological　samples　and　is　suitable　for　use　in　the　mass　production　of　miniaturized　devices,　thus　opening　new　opportunities　for　protein　diagnostics　and　biosecurity.　©　2017　American　Chemical　Society.