title: DNA aptamer-based sandwich microfluidic assays for dual quantification and multi-glycan profiling of cancer biomarkers.
abstract: Two novel sandwich-based immunoassays for prostate cancer (PCa) diagnosis are reported, in which the primary antibody for capture is replaced by a DNA aptamer. The assays, which can be performed in parallel, were developed in a microfluidic device and tested for the detection of free Prostate Specific Antigen (fPSA). A secondary antibody (Aptamer-Antibody Assay) or a lectin (Aptamer-Lectin Assay) is used to quantify, by chemiluminescence, both the amount of fPSA and its glycosylation levels. The use of aptamers enables a more reliable, selective and controlled sensing of the analyte. The dual approach provides sensitive detection of fPSA along with selective fPSA glycoprofiling, which is of significant importance in the diagnosis and prognosis of PCa, as tumor progression is associated with changes in fPSA glycosylation. With these approaches, we can potentially detect 0.5 ng/mL of fPSA and 3 ng/mL of glycosylated fPSA using Sambucus nigra (SNA) lectin, both within the relevant clinical range. The approach can be applied to a wide range of biomarkers, thus providing a good alternative to standard antibody-based immunoassays with significant impact in medical diagnosis and prognosis.
authors.affiliation: Department of Electronic & Electrical Engineering, University of Bath, Bath BA2 7AY, United Kingdom. Electronic address: P.Jolly@bath.ac.uk.
authors.forename: Pawan
authors.initials: P
authors.lastname: Jolly
authors.affiliation: Department of Glycobiotechnology, Center for Glycomics, Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava 84105, Slovakia. Electronic address: Pavel.Damborsky@savba.sk.
authors.forename: Pavel
authors.initials: P
authors.lastname: Damborsky
authors.affiliation: INESC-MN - Microsystems and Nanotechnologies, R. Alves Redol 9, 1000-029 Lisboa, Portugal. Electronic address: Nsrinivasan@inesc-mn.pt.
authors.forename: Narayanan
authors.initials: N
authors.lastname: Madaboosi
authors.affiliation: INESC-MN - Microsystems and Nanotechnologies, R. Alves Redol 9, 1000-029 Lisboa, Portugal; Department of Bioengineering, Instituto Superior Técnico, University of Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal. Electronic address: Ruben.Soares@tecnico.ulisboa.pt.
authors.forename: Ruben R G
authors.initials: RR
authors.lastname: Soares
authors.affiliation: INESC-MN - Microsystems and Nanotechnologies, R. Alves Redol 9, 1000-029 Lisboa, Portugal. Electronic address: vchu@inesc-mn.pt.
authors.forename: Virginia
authors.initials: V
authors.lastname: Chu
authors.affiliation: INESC-MN - Microsystems and Nanotechnologies, R. Alves Redol 9, 1000-029 Lisboa, Portugal; Department of Bioengineering, Instituto Superior Técnico, University of Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal. Electronic address: joao.conde@tecnico.ulisboa.pt.
authors.forename: João P
authors.initials: JP
authors.lastname: Conde
authors.affiliation: Department of Glycobiotechnology, Center for Glycomics, Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava 84105, Slovakia. Electronic address: Jaroslav.Katrlik@savba.sk.
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Katrlik
authors.affiliation: Department of Electronic & Electrical Engineering, University of Bath, Bath BA2 7AY, United Kingdom. Electronic address: P.Estrela@bath.ac.uk.
authors.forename: Pedro
authors.initials: P
authors.lastname: Estrela
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000302:isolation & purification*; D015374:Biosensing Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008297:Male; D044085:Microfluidics / Q000379:methods; D011134:Polysaccharides / Q000302:isolation & purification; D011379:Prognosis; D017430:Prostate-Specific Antigen / Q000302:isolation & purification*; D011471:Prostatic Neoplasms / Q000175:diagnosis* / Q000473:pathology
chemical_list: D052157:Aptamers, Nucleotide; D014408:Biomarkers, Tumor; D011134:Polysaccharides; D017430:Prostate-Specific Antigen
keywords: DNA aptamer; ELISA; Glycoprofiling; Microfluidics; Prostate cancer; Sandwich assay