title: Biofunctionalized magnetic nanospheres-based cell sorting strategy for efficient isolation, detection and subtype analyses of heterogeneous circulating hepatocellular carcinoma cells.
abstract: Hepatocellular carcinoma (HCC) is an awful threat to human health. Early-stage HCC may be detected by isolation of circulating tumor cells (CTCs) from peripheral blood samples, which is beneficial to the diagnosis and therapy. However, the extreme rarity and high heterogeneity of HCC CTCs have been restricting the relevant research. To achieve an efficient isolation, reliable detection and subtype analyses of heterogeneous HCC CTCs, herein, we present a cell sorting strategy based on anti-CD45 antibody-modified magnetic nanospheres. By this strategy, leukocyte depletion efficiency was up to 99.9% within 30min in mimic clinical samples, and the purity of the spiked HCC cells was improved 265-317-fold. Besides, the isolated HCC cells remained viable at 92.3% and could be directly recultured. Moreover, coupling the convenient, fast and effective cell sorting strategy with specific ICC identification via biomarkers AFP and GPC3, HCC CTCs were detectable in peripheral blood samples, showing the potential for HCC CTC detection in clinic. Notably, this immunomagnetic cell sorting strategy enabled isolating more heterogeneous HCC cells compared with the established EpCAM-based methods, and further achieved characterization of three different CTC subtypes from one clinical HCC blood sample, which may assist clinical HCC analyses such as prognosis or personalized treatment.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China.
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China.
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Wu
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China.
authors.forename: Zhi-Ling
authors.initials: ZL
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China.
authors.forename: Jiao
authors.initials: J
authors.lastname: Hu
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China.
authors.forename: Man
authors.initials: M
authors.lastname: Tang
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China; Hubei Cancer Hospital, Wuhan 430079, PR China.
authors.forename: Chu-Bo
authors.initials: CB
authors.lastname: Qi
authors.affiliation: Renmin Hospital of Wuhan University, Wuhan 430060, PR China.
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, The Institute for Advanced Studies, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China. Electronic address: dwpang@whu.edu.cn.
authors.forename: Dai-Wen
authors.initials: DW
authors.lastname: Pang
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000473:pathology*; D002469:Cell Separation / Q000379:methods*; D056945:Hep G2 Cells; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D019169:Jurkat Cells; D047589:Leukocyte Reduction Procedures / Q000379:methods; D008113:Liver Neoplasms / Q000097:blood / Q000473:pathology*; D054118:Nanospheres / Q000737:chemistry; D009360:Neoplastic Cells, Circulating / Q000473:pathology*
chemical_list: D055910:Antibodies, Immobilized
keywords: Circulating tumor cell; Hepatocellular carcinoma; Immunomagnetic; Isolation; Nanosphere