title: Detection of carcinoembryonic antigen using single-domain or full-size antibodies stained with quantum dot conjugates.
abstract: Compact single-domain antibodies (sdAbs) are nearly 13 times smaller than full-size monoclonal antibodies (mAbs) and have a number of advantages for biotechnological applications, such as small size, high specificity, solubility, stability, and great refolding capacity. Carcinoembryonic antigen (CEA) is a tumor-associated glycoprotein expressed in a variety of cancers. Detection of CEA on the tumor cell surface may be carried out using anti-CEA antibodies and conventional fluorescent dyes. Semiconductor quantum dots (QDs) are brighter and more photostable than organic dyes; they provide the possibility for labeling of different recognition molecules with QDs of different colors but excitable with the same wavelength of excitation. In this study, the abilities for specific detection of CEA expressed by tumor cells with anti-CEA sdAbs biotinylated in vitro and in vivo, as well as with anti-CEA mAbs biotinylated in vitro, were compared using flow cytometry and the conjugates of streptavidin with QDs (SA-QDs). The results demonstrated that either in vitro or in vivo biotinylated anti-CEA sdAbs are more sensitive for cell staining compared to biotinylated anti-CEA mAbs. The data also show that simultaneous use of biotinylated sdAbs with highly fluorescent SA-QDs can considerably improve the sensitivity of detection of CEA on tumor cell surfaces.
authors.affiliation: Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Rousserie
authors.affiliation: Laboratory of Nano-Bioengineering, National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe Shosse, 115409 Moscow, Russian Federation.
authors.forename: Regina
authors.initials: R
authors.lastname: Grinevich
authors.affiliation: Laboratory of Nano-Bioengineering, National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe Shosse, 115409 Moscow, Russian Federation.
authors.forename: Kristina
authors.initials: K
authors.lastname: Brazhnik
authors.affiliation: INSERM U1068 and CNRS UMR7258, Centre de Recherche en Cancérologie de Marseille, Institut Paoli-Calmettes and Université Aix-Marseille, 13009 Marseille, France.
authors.forename: Klervi
authors.initials: K
authors.lastname: Even-Desrumeaux
authors.affiliation: Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Reveil
authors.affiliation: Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Tabary
authors.affiliation: INSERM U1068 and CNRS UMR7258, Centre de Recherche en Cancérologie de Marseille, Institut Paoli-Calmettes and Université Aix-Marseille, 13009 Marseille, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Chames
authors.affiliation: INSERM U1068 and CNRS UMR7258, Centre de Recherche en Cancérologie de Marseille, Institut Paoli-Calmettes and Université Aix-Marseille, 13009 Marseille, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Baty
authors.affiliation: Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France.
authors.forename: Jacques H M
authors.initials: JH
authors.lastname: Cohen
authors.affiliation: Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France; Laboratory of Nano-Bioengineering, National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe Shosse, 115409 Moscow, Russian Federation. Electronic address: igor.nabiev@gmail.com.
authors.forename: Igor
authors.initials: I
authors.lastname: Nabiev
authors.affiliation: Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France; Laboratory of Nano-Bioengineering, National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe Shosse, 115409 Moscow, Russian Federation. Electronic address: alyona.sukhanova@univ-reims.fr.
authors.forename: Alyona
authors.initials: A
authors.lastname: Sukhanova
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D019921:Biotinylation; D002272:Carcinoembryonic Antigen / Q000032:analysis*; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D045663:Quantum Dots / Q000737:chemistry*; D061905:Single-Domain Antibodies / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D061905:Single-Domain Antibodies
keywords: Biotinylation; Carcinoembryonic antigen (CEA); Quantum dots; Single-domain antibodies (sdAbs)