Journal
ACS SENSORS
Volume 2, Issue 9, Pages 1267-1271Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.7b00495
Keywords
nanobody; apolipoprotein E; point-of-care testing; colorimetric; immunosensor
Funding
- National Key Scientific Instrument and Equipment Development Project of China [21627809]
- National Natural Science Foundation of China [21375047, 21575137, 21575050, 21601064]
- Special Foundation for Taishan Scholar Professorship of Shandong Province [ts20130937]
- Science and Technology Development Plan of Shandong Province [2014GSF120004]
- Graduate Innovation Foundation of UJN [YCXB15004]
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Alzheimer's disease (AD) biomarkers can reflect the neurochemical indicators used to estimate the risk in clinical nephrology. Apolipoprotein E (ApoE) is an early biomarker for AD in clinical diagnosis. In this research, through bactrian camel immunization, lymphocyte isolation, RNA extraction, and library construction, ApoE-specific Nbs with high affinity were successfully separated from an immune phage display nanobody library. Herein, a colorimetric immunosensor was developed for the point-of-care testing of ApoE by layer-by-layer nanoassembly techniques and novel nanobodies (Nbs). Using highly oriented Nbs as the capture and detection antibodies, an on-site immunosensor was developed by detecting the mean gray value of fade color due to the glutaraldehydeP3-aminopropyltrimethoxysilane oxidation by H2O2. The detection limit of AopE is 0.42 pg/mL, and the clinical analysis achieves a good performance. The novel easily operated immunosensor may have potential application in the clinical diagnosis and real-time monitoring for AD.
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