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dc.contributor.author
Seia, Marco Abel
dc.contributor.author
Stege, Patricia Wanda
dc.contributor.author
Pereira, Sirley Vanesa
dc.contributor.author
de Vito, Irma Esther
dc.contributor.author
Raba, Julio
dc.contributor.author
Messina, Germán Alejandro
dc.date.available
2016-05-17T15:18:52Z
dc.date.issued
2014-06
dc.identifier.citation
Seia, Marco Abel; Stege, Patricia Wanda; Pereira, Sirley Vanesa; de Vito, Irma Esther; Raba, Julio; et al.; Silica nanoparticle-based microfluidic immunosensor with laser-induced fluorescence detection for the quantification of immunoreactive trypsin; Elsevier; Analytical Biochemistry; 463; 6-2014; 31-37
dc.identifier.issn
0003-2697
dc.identifier.uri
http://hdl.handle.net/11336/5710
dc.description.abstract
The purpose of this study was to develop a silica nanoparticle-based immunosensor with laser-induced fluorescence (LIF) as a detection system. The proposed device was applied to quantify the immunoreactive trypsin (IRT) in cystic fibrosis (CF) newborn screening. A new ultrasonic procedure was used to extract the IRT from blood spot samples collected on filter papers. After extraction, the IRT reacted immunologically with anti-IRT monoclonal antibodies immobilized on a microfluidic glass chip modified with 3-aminopropyl functionalized silica nanoparticles (APSN–APTES-modified glass chips). The bounded IRT was quantified by horseradish peroxidase (HRP)-conjugated anti-IRT antibody (anti-IRT–Ab) using 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) as enzymatic mediator. The HRP catalyzed the oxidation of nonfluorescent ADHP to highly fluorescent resorufin, which was measured by LIF detector, using excitation lambda at 561 nm and emission at 585 nm. The detection limits (LODs) calculated for LIF detection and for a commercial enzyme-linked immunosorbent assay (ELISA) test kit were 0.87 and 4.2 ng ml−1, respectively. The within- and between-assay variation coefficients for the LIF detection procedure were below 6.5%. The blood spot samples collected on filter papers were analyzed with the proposed method, and the results were compared with those of the reference ELISA method, demonstrating a potential usefulness for the clinical assessment of IRT during the early neonatal period.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.subject
Newborn Screening
dc.subject
Immunoreactive Trypsin
dc.subject
Microfluidic Chip
dc.subject
Silica Nanoparticles
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Ultrasonic Procedure
dc.subject
Lif
dc.subject.classification
Química Analítica
dc.subject.classification
Ciencias Químicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Silica nanoparticle-based microfluidic immunosensor with laser-induced fluorescence detection for the quantification of immunoreactive trypsin
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2016-05-16T20:18:59Z
dc.journal.volume
463
dc.journal.pagination
31-37
dc.journal.pais
Países Bajos
dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: Seia, Marco Abel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Química de San Luis; Argentina
dc.description.fil
Fil: Stege, Patricia Wanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Química de San Luis; Argentina
dc.description.fil
Fil: Pereira, Sirley Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Química de San Luis; Argentina
dc.description.fil
Fil: de Vito, Irma Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Química de San Luis; Argentina
dc.description.fil
Fil: Raba, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Química de San Luis; Argentina
dc.description.fil
Fil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico San Luis. Instituto de Química de San Luis; Argentina
dc.journal.title
Analytical Biochemistry
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0003269714002759
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.ab.2014.06.016
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ab.2014.06.016
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