title: Bioconjugated and cross-linked bionanostructures for bifunctional immunohistochemical labeling.
abstract: The present study describes the development and use of a new bioconjugate combining targeted quantum dot labeling with an immunoperoxidase method and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1). The new bioconjugate is designed for the examination of samples both under fluorescent and bright-field microscopy at the same time. For this purpose chlorobis(2-2'-bipyridyl) methacryloyl tyrosine-ruthenium(II) and bis (2-2'-bipyridyl) methacryloyltyrosine-methacryloyltryptophan-ruthenium (II) photosensitive monomers and photosensitive poly(Bis (2-2'-bipyridyl)) methacryloyltyrosine-methacryloyltryptophan-ruthenium(II) were synthesized and characterized. The anti-SOD1 antibody and horseradish peroxidase (HRD) conjugated quantum dots were prepared by using this polymer. The anti-SOD1 antibody and HRD conjugated quantum dots were used in labeling and imaging of SOD1 in rat liver sections. Quantum dot particles were observed as a bright fluorescence in their specific binding locations inside the hepatocytes. The HRD-diaminobenzidine reaction product was observed as brown-colored particles at the same locations under bright-field microscopy. Structural details of the tissue sections could be examined at the same time. The conjugation protocol is simple; the bioconjugate is applicable for efficient cell labeling and can be adapted for imaging of other targets in different tissues. Also, the prepared nanobioconjugates have mechanic stability and can be used for a long period.
authors.affiliation: Department of Chemistry, Anadolu University, Eskişehir, Turkey. rsay@anadolu.edu.tr
authors.forename: Rıdvan
authors.initials: R
authors.lastname: Say
authors.forename: Gözde Aydoğan
authors.initials: GA
authors.lastname: Kılıç
authors.forename: Ayça Atılır
authors.initials: AA
authors.lastname: Ozcan
authors.forename: Deniz
authors.initials: D
authors.lastname: Hür
authors.forename: Filiz
authors.initials: F
authors.lastname: Yılmaz
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
authors.forename: Arzu
authors.initials: A
authors.lastname: Ersöz
mesh_terms: D000818:Animals; D006735:Horseradish Peroxidase / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods*; D008099:Liver / Q000201:enzymology / Q000473:pathology; D008853:Microscopy / Q000379:methods; D045663:Quantum Dots; D051381:Rats; D013194:Staining and Labeling / Q000379:methods*; D013482:Superoxide Dismutase / Q000032:analysis; D000072105:Superoxide Dismutase-1
chemical_list: D006735:Horseradish Peroxidase; C000606292:Sod1 protein, rat; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1