Abstract:ELISA (enzyme-linked immunosorbent assay), a highly sensitive and powerful molecular detection tool, has been constructed based on antibodies, as its name denotes. However, it is not so easy to prepare antibodies for any target molecule and there are still problems in their cost and stability. In this study, peptide-based ELISA-like systems (pep-ELISA) were first constructed and shown to be effective. In particular, if the target is an enzyme, such as cathepsin E, that can generate a fluorescent product, the c… Show more
“…To address these challenges, a microfluidic approach can be used to reduce the possible non-specific interactions. Our sensor provided an improvement of sensitivity of serum CatE by at least three orders of magnitude compared to a conventional ELISA system using a similar peptide aptamer 19 . Given this observation and with implementation of further improvements in the sampling process, our peptide-modified SWCNT FET biosensor could offer an attractive approach to detect CatE in a real clinical sample with higher selectivity and the desired sensitivity.…”
Section: Resultsmentioning
confidence: 96%
“…Our device exhibited good selectivity (no response for bovine serum albumin (BSA) and cathepsin K (CatK)) and a low detection limit. The sensitivity of this device could improve the limit of detection of CatE in human serum by at least three orders of magnitude compared with a conventional ELISA-based system using a similar peptide aptamer 19 . Figure 1 Schematic of the concept.…”
Biosensors employing single-walled carbon nanotube field-effect transistors (SWCNT FETs) offer ultimate sensitivity. However, besides the sensitivity, a high selectivity is critically important to distinguish the true signal from interference signals in a non-controlled environment. This work presents the first demonstration of the successful integration of a novel peptide aptamer with a liquid-gated SWCNT FET to achieve highly sensitive and specific detection of Cathepsin E (CatE), a useful prognostic biomarker for cancer diagnosis. Novel peptide aptamers that specifically recognize CatE are engineered by systemic in vitro evolution. The SWCNTs were firstly grown using the thermal chemical vapor deposition (CVD) method and then were employed as a channel to fabricate a SWCNT FET device. Next, the SWCNTs were functionalized by noncovalent immobilization of the peptide aptamer using 1-pyrenebutanoic acid succinimidyl ester (PBASE) linker. The resulting FET sensors exhibited a high selectivity (no response to bovine serum albumin and cathepsin K) and label-free detection of CatE at unprecedentedly low concentrations in both phosphate-buffered saline (2.3 pM) and human serum (0.23 nM). Our results highlight the use of peptide aptamer-modified SWCNT FET sensors as a promising platform for near-patient testing and point-of-care testing applications.
“…To address these challenges, a microfluidic approach can be used to reduce the possible non-specific interactions. Our sensor provided an improvement of sensitivity of serum CatE by at least three orders of magnitude compared to a conventional ELISA system using a similar peptide aptamer 19 . Given this observation and with implementation of further improvements in the sampling process, our peptide-modified SWCNT FET biosensor could offer an attractive approach to detect CatE in a real clinical sample with higher selectivity and the desired sensitivity.…”
Section: Resultsmentioning
confidence: 96%
“…Our device exhibited good selectivity (no response for bovine serum albumin (BSA) and cathepsin K (CatK)) and a low detection limit. The sensitivity of this device could improve the limit of detection of CatE in human serum by at least three orders of magnitude compared with a conventional ELISA-based system using a similar peptide aptamer 19 . Figure 1 Schematic of the concept.…”
Biosensors employing single-walled carbon nanotube field-effect transistors (SWCNT FETs) offer ultimate sensitivity. However, besides the sensitivity, a high selectivity is critically important to distinguish the true signal from interference signals in a non-controlled environment. This work presents the first demonstration of the successful integration of a novel peptide aptamer with a liquid-gated SWCNT FET to achieve highly sensitive and specific detection of Cathepsin E (CatE), a useful prognostic biomarker for cancer diagnosis. Novel peptide aptamers that specifically recognize CatE are engineered by systemic in vitro evolution. The SWCNTs were firstly grown using the thermal chemical vapor deposition (CVD) method and then were employed as a channel to fabricate a SWCNT FET device. Next, the SWCNTs were functionalized by noncovalent immobilization of the peptide aptamer using 1-pyrenebutanoic acid succinimidyl ester (PBASE) linker. The resulting FET sensors exhibited a high selectivity (no response to bovine serum albumin and cathepsin K) and label-free detection of CatE at unprecedentedly low concentrations in both phosphate-buffered saline (2.3 pM) and human serum (0.23 nM). Our results highlight the use of peptide aptamer-modified SWCNT FET sensors as a promising platform for near-patient testing and point-of-care testing applications.
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