{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T19:46:46Z","timestamp":1773690406042,"version":"3.50.1"},"reference-count":55,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2009,11,19]],"date-time":"2009-11-19T00:00:00Z","timestamp":1258588800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>The early detection of HER2 (human epidermal growth factor receptor 2) status in breast cancer patients is very important for the effective implementation of anti-HER2 antibody therapy. Recently, HER2 detections using antibody conjugated quantum dots (QDs) have attracted much attention. QDs are a new class of fluorescent materials that have superior properties such as high brightness, high resistance to photo-bleaching, and multi-colored emission by a single-light source excitation. In this study, we synthesized three types of anti-HER2 antibody conjugated QDs (HER2Ab-QDs) using different coupling agents (EDC\/sulfo-NHS, iminothiolane\/sulfo-SMCC, and sulfo-SMCC). As water-soluble QDs for the conjugation of antibody, we used glutathione coated CdSe\/CdZnS QDs (GSH-QDs) with fluorescence quantum yields of 0.23~0.39 in aqueous solution. Dispersibility, hydrodynamic size, and apparent molecular weights of the GSH-QDs and HER2Ab-QDs were characterized by using dynamic light scattering, fluorescence correlation spectroscopy, atomic force microscope, and size-exclusion HPLC. Fluorescence imaging of HER2 overexpressing cells (KPL-4 human breast cancer cell line) was performed by using HER2Ab-QDs as fluorescent probes. We found that the HER2Ab-QD prepared by using SMCC coupling with partially reduced antibody is a most effective probe for the detection of HER2 expression in KPL-4 cells. We have also studied the size dependency of HER2Ab-QDs (with green, orange, and red emission) on the fluorescence image of KPL-4 cells.<\/jats:p>","DOI":"10.3390\/s91109332","type":"journal-article","created":{"date-parts":[[2009,11,19]],"date-time":"2009-11-19T12:39:06Z","timestamp":1258634346000},"page":"9332-9354","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":70,"title":["Synthesis and Characterization of Anti-HER2 Antibody Conjugated CdSe\/CdZnS Quantum Dots for Fluorescence Imaging of Breast Cancer Cells"],"prefix":"10.3390","volume":"9","author":[{"given":"Dhermendra K.","family":"Tiwari","sequence":"first","affiliation":[{"name":"WPI Immunology Frontier Research Center, Osaka University, Yamada-oka 1-3, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shin-Ichi","family":"Tanaka","sequence":"additional","affiliation":[{"name":"Graduate School of Frontier Biosciences, Osaka University, Yamada-oka 2-1, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yasushi","family":"Inouye","sequence":"additional","affiliation":[{"name":"Graduate School of Frontier Biosciences, Osaka University, Yamada-oka 2-1, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keiko","family":"Yoshizawa","sequence":"additional","affiliation":[{"name":"WPI Immunology Frontier Research Center, Osaka University, Yamada-oka 1-3, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomonobu M.","family":"Watanabe","sequence":"additional","affiliation":[{"name":"WPI Immunology Frontier Research Center, Osaka University, Yamada-oka 1-3, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takashi","family":"Jin","sequence":"additional","affiliation":[{"name":"WPI Immunology Frontier Research Center, Osaka University, Yamada-oka 1-3, Suita, Osaka 565-0871, Japan"},{"name":"Graduate School of Frontier Biosciences, Osaka University, Yamada-oka 2-1, Suita, Osaka 565-0871, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2009,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1126\/science.3798106","article-title":"Human breast cancer: correlation of relapse and survival with amplification of the HER-2\/neu oncogene","volume":"235","author":"Slamon","year":"1987","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1126\/science.2470152","article-title":"Studies of the HER-2\/neu proto-oncogene in human breast and ovarian cancer","volume":"244","author":"Slamon","year":"1989","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2639","DOI":"10.1200\/JCO.1999.17.9.2639","article-title":"Multinational study of the efficacy and safety of humanized anti-HER2 monoclonal antibody in women who have HER2-overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease","volume":"17","author":"Cobleigh","year":"1999","journal-title":"J. Clin. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1056\/NEJMra043186","article-title":"Trastuzumab-mechanism of action and use in clinical practice","volume":"357","author":"Haudis","year":"2007","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1200\/JCO.2006.06.4220","article-title":"Cost effectiveness of adjuvant trastuzumab in human epidermal growth factor receptor-2 positive breast cancer","volume":"25","author":"Liberato","year":"2007","journal-title":"J. Clin. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/modpathol.3880007","article-title":"Determination of Her2-\/Neu status in breast carcinoma: comparative analysis of immunohistochemistry and fluorescent in situ hybridization","volume":"13","author":"Jimenez","year":"2000","journal-title":"Mod. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1111\/j.1365-2559.2006.02513.x","article-title":"Quantification of immunohistochemistry-issues concerning methods, utility and semiquantitative assessment II","volume":"49","author":"Taylor","year":"2006","journal-title":"Histopathology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1111\/j.1365-2559.2007.02897.x","article-title":"Predictive markers in breast cancer-the present","volume":"52","author":"Payne","year":"2008","journal-title":"Histopathology"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1002\/path.1523","article-title":"Comparative multi-methodological measurement of ERBB2 status in breast cancer","volume":"202","author":"Ginestier","year":"2004","journal-title":"J. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1972","DOI":"10.1001\/jama.291.16.1972","article-title":"HER-2 testing in breast cancer using parallel tissue-based methods","volume":"29","author":"Yaziji","year":"2004","journal-title":"J. Am. Med. Assn."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/nbt764","article-title":"Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots","volume":"21","author":"Wu","year":"2003","journal-title":"Nat. Biotechnol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.bbrc.2006.09.159","article-title":"Reduction in nonfluorescence state of quantum dots on an immunofluorescence staining","volume":"351","author":"Watanabe","year":"2006","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1138","DOI":"10.1158\/0008-5472.CAN-06-1185","article-title":"In vivo real-time tracking of single quantum dots conjugated with monoclonal anti-HER2 antibody in tumors of mice","volume":"67","author":"Tada","year":"2007","journal-title":"Cancer Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4109","DOI":"10.1529\/biophysj.106.094649","article-title":"Stepwise movements in visible transport of HER2 by motor proteins in living cells","volume":"92","author":"Watanabe","year":"2007","journal-title":"Biophys. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2871","DOI":"10.1021\/ja067184n","article-title":"Forming biocompatible and nonaggregated nanocrystals in water using amphiphilic polymers","volume":"129","author":"Yu","year":"2007","journal-title":"J. Am. Chem. Soc."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1038\/nprot.2007.107","article-title":"Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry","volume":"2","author":"Xing","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s12282-008-0037-0","article-title":"In vivo single molecular imaging and sentinel node navigation by nanotechnology for molecular targeting drug-delivery systems and tailor-made medicine","volume":"15","author":"Takeda","year":"2008","journal-title":"Breast Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2178","DOI":"10.1002\/ijc.23320","article-title":"Quantitation of HER2 and telomerase biomarkers in solid tumors with IgY antibodies and nanocrystal detection","volume":"122","author":"Xiao","year":"2008","journal-title":"Int. J. Cancer"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2912","DOI":"10.1016\/j.biomaterials.2009.02.010","article-title":"Quantum dots-based immunofluorescence technology for the quantitative determination of HER2 expression in breast cancer","volume":"30","author":"Chen","year":"2009","journal-title":"Biomaterials"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1002\/smll.200800714","article-title":"Single chain epidermal growth factor antibody conjugated nanoparticles for in vivo tumor targeting and imaing","volume":"5","author":"Yang","year":"2009","journal-title":"Small"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"021004","DOI":"10.1117\/1.3122775","article-title":"Fluorescent immunolabeling of cancer cells by quantum dots and antibody scFv fragment","volume":"14","author":"Zdobnova","year":"2009","journal-title":"J. Biomed. Opt."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1307","DOI":"10.1373\/clinchem.2008.120576","article-title":"Cell lines candidate reference materials for quality control of ERBB2 amplification and expression assays in breast cancer","volume":"55","author":"Xiao","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_23","unstructured":"Hermanson, G.T. (1996). Bioconjugate Techniques, Academic Press."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"14530","DOI":"10.1021\/ja073790m","article-title":"Compact cystein-coated CdSe(ZnCdS) quantum dots for in vivo applications","volume":"129","author":"Liu","year":"2007","journal-title":"J. Am. Chem. Soc."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1021\/jp9530562","article-title":"Synthesis and characterization of strongly luminescent ZnS-capped CdSe nanocrystals","volume":"100","author":"Hines","year":"1996","journal-title":"J. Phys. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8706","DOI":"10.1021\/ja00072a025","article-title":"Synthesis and characterization of nearly monodisperse CdE (E = S, Se, Te) semiconductor nanocrystallites","volume":"115","author":"Murray","year":"1993","journal-title":"J. Am. Chem. Soc."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"7019","DOI":"10.1021\/ja970754m","article-title":"Epitaxial growth of highly luminescent CdSe\/CdS core\/shell nanocrystals with photostability and electronic accessibility","volume":"119","author":"Peng","year":"1997","journal-title":"J. Am. Chem. Soc."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9463","DOI":"10.1021\/jp971091y","article-title":"(CdSe)ZnS coe-shell quantum dots: synthesis and characterization of a size series of highly luminescent nanocrystallites","volume":"101","author":"Dabbousi","year":"1997","journal-title":"J. Phys. Chem. B"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2049","DOI":"10.1021\/ja017002j","article-title":"Control of photoluminescence properties of CdSe nanocrystals in growth","volume":"124","author":"Qu","year":"2002","journal-title":"J. Am. Chem. Soc."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1676","DOI":"10.1002\/anie.200704794","article-title":"Optical detection of glucose and acetylcholine esterase inhibitors by H2O2-sensitive CdSe\/ZnS quantum dots","volume":"47","author":"Gill","year":"2008","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2044","DOI":"10.3390\/ijms9102044","article-title":"Preparation and characterization of highly fluorescent, glutathione-coated near infrared quantum dots for in vivo fluorescence imaging","volume":"9","author":"Jin","year":"2008","journal-title":"Int. J. Mol. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Jin, T., Yoshioka, Y., Fujii, F., Komai, Y., Seki, J., and Seiyama, A (2008). Gd3+-functionalized near-infrared quantum dots for in vivo dual modal (fluorescence\/magnetic resonance) imaging. Chem. Commun., 5764\u20135766.","DOI":"10.1039\/b812302k"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"10149","DOI":"10.1158\/0008-5472.CAN-05-1781","article-title":"Nonivasive in vivo detection of glutathione metabolism in tumors","volume":"65","author":"Thelwall","year":"2005","journal-title":"Cancer. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.neures.2006.01.002","article-title":"Contribution of reduced and oxidized glutathione to signals detected by magnetic resonance spectroscopy as indicators of local brain redox state","volume":"55","author":"Satoh","year":"2006","journal-title":"Neurosci. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1096\/fasebj.1.3.2887478","article-title":"Glutathione, a first defense against cadmium toxicity","volume":"1","author":"Singhal","year":"1987","journal-title":"FASEB J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1038\/nmeth.1206","article-title":"Monovalent, reduced-size quantum dots for imaging receptors on living cells","volume":"5","author":"Howarth","year":"2008","journal-title":"Nat. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1274","DOI":"10.1021\/ja076069p","article-title":"Compact biocompatible quantum dots functionalized for cellular imaging","volume":"130","author":"Liu","year":"2008","journal-title":"J. Am. Chem. Soc."},{"key":"ref_38","first-page":"3854s","article-title":"Effect of molecular size of pegylated peptide on the pharmacokinetics and tumor targeting in lymphoma-nearing mice","volume":"9","author":"Denardo","year":"2003","journal-title":"Clin. Can. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1165","DOI":"10.1038\/nbt1340","article-title":"Renal clearence of quantum dots","volume":"25","author":"Choi","year":"2007","journal-title":"Nat. Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2829","DOI":"10.1039\/b503178h","article-title":"Calixarene-coated water-soluble CdSe-ZnS semiconductor quantum dots that are highly fluorescent and stable in aqueous solution","volume":"22","author":"Jin","year":"2005","journal-title":"Chem. Commun."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4300","DOI":"10.1039\/b506608e","article-title":"Amphiphilic p-sulfonatocalix[4]arene-coated CdSe\/ZnS quantum dots for the optical detection of the neurotransmitter acetylcholine","volume":"34","author":"Jin","year":"2005","journal-title":"Chem. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"9288","DOI":"10.1021\/ja062021k","article-title":"Control of the optical properties of quantum dots by surface coating with calix[n]arene carboxylic acids","volume":"128","author":"Jin","year":"2006","journal-title":"J. Am. Chem. Soc."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1870","DOI":"10.1039\/b617115j","article-title":"Fluorescence correlation spectroscopy using quantum dots: advances, challenges and opportunities","volume":"9","author":"Heuff","year":"2007","journal-title":"Phys. Chem. Chem. Phys."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"473","DOI":"10.2174\/138620707781996466","article-title":"Preparation and characterization of thiacalix[4]arene coated water-soluble CdSe\/ZnS quantum dots as a fluorescent probe for Cu2+ ions","volume":"10","author":"Jin","year":"2007","journal-title":"Comb. Chem. High Throughput Scr."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1711","DOI":"10.1021\/nl0707003","article-title":"Simultaneous multicolor imaging of five different lymphatic basins using quantum dots","volume":"7","author":"Kobayashi","year":"2007","journal-title":"Nano Lett."},{"key":"ref_46","first-page":"747","article-title":"Crystal structure of the aequorea Victoria green fluorescent protein","volume":"273","author":"Cubitt","year":"1986","journal-title":"Science"},{"key":"ref_47","first-page":"1392","article-title":"Three-dimensional structure of an antigen-antibody complex at 2.5 \u00c5 resulution","volume":"273","author":"Amit","year":"1996","journal-title":"Science"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/S1389-0344(02)00046-1","article-title":"Investigation specific antigen\/antibody binding with the atomic force microscope","volume":"19","author":"Ouerghi","year":"2002","journal-title":"Biomol. Eng."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"4103","DOI":"10.1021\/ja0058334","article-title":"Hydroxylated quantum dots as luminescent probes for in situ hybridization","volume":"123","author":"Pathak","year":"2001","journal-title":"J. Am. Chem. Soc."},{"key":"ref_50","first-page":"10","article-title":"Capillary electrophoresis for the characterization of quantum dots after non-selective or selective bioconjugation with antibodies for immunoassay","volume":"6","author":"Pereira","year":"2008","journal-title":"J. Nanotechnol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"5268","DOI":"10.1091\/mbc.e04-07-0591","article-title":"Endocytosis and sorting of ErbB2 and the site of action of cancer therapeutics trastuzumab and geldanamycin","volume":"15","author":"Austin","year":"2004","journal-title":"Mol. Biol. Cell."},{"key":"ref_52","first-page":"3275","article-title":"Specific tumoricidal activity of a secreted proaptotic protein consisting of HER2 antibody and constutively active caspase-3","volume":"63","author":"Jia","year":"2003","journal-title":"Cancer Res."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Rigler, R., and Elson, E.S. (2001). Fluorescence correlation spectroscopy, Springer-Verlag.","DOI":"10.1007\/978-3-642-59542-4"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1038\/sj.bjc.6690114","article-title":"Isolation and characterization of a new human breat cancer cell line, KPL-4, expressing the Erb B family receptors and interleukin-6","volume":"79","author":"Kurebayashi","year":"1998","journal-title":"Br. J. Cancer"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s00280-001-0401-7","article-title":"Antitumor activity of combinations of anti-HER-2 antibody trastuzumab and oral fluoropyrimidines capecitabine\/5\u2032-dFUrd in human breast cancer models","volume":"49","author":"Ouchi","year":"2002","journal-title":"Cancer Chemother. Pharmacol."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/9\/11\/9332\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T22:11:43Z","timestamp":1760220703000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/9\/11\/9332"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2009,11,19]]},"references-count":55,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2009,11]]}},"alternative-id":["s91109332"],"URL":"https:\/\/doi.org\/10.3390\/s91109332","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2009,11,19]]}}}