title: Site-selective in situ growth of fluorescent polymer-antibody conjugates with enhanced antigen detection by signal amplification.
abstract: This paper reports a new and general in situ methodology to grow fluorescent polymer conjugates from the interchain disulfide bridging sites of a monoclonal antibody. Atom transfer radical polymerization (ATRP) initiators were attached to a monoclonal antibody at its interchain disulfide bridging sites by disulfide re-bridging to yield a macroinitiator. Subsequent in situ ATRP of PEG-like monomers with dye-functionalized monomers from the macroinitiator formed antibody-polymer-dye conjugates with site-selectivity and tunable dye-to-antibody ratios. Notably, these conjugates can amplify antigen detection signal by reducing label-density dependent fluorescence quenching and by increasing dye-to-antibody ratios. The method developed may be applicable to a variety of antibodies, dyes and drugs to create a number of antibody-polymer-dye/drug conjugates for advanced diagnosis and therapy of diseases.
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Libin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Wenguo
authors.initials: W
authors.lastname: Zhao
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Xinyu
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
authors.forename: Guilin
authors.initials: G
authors.lastname: Wang
authors.affiliation: Sino Biological, Inc., 14 Zhonghe St. BDA, Beijing 100176, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Sino Biological, Inc., 14 Zhonghe St. BDA, Beijing 100176, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Li
authors.affiliation: Sino Biological, Inc., 14 Zhonghe St. BDA, Beijing 100176, China.
authors.forename: Liangzhi
authors.initials: L
authors.lastname: Xie
authors.affiliation: Protein Chemistry Facility, School of Biological Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Protein Chemistry Facility, School of Biological Sciences, Tsinghua University, Beijing 100084, China.
authors.forename: Haiteng
authors.initials: H
authors.lastname: Deng
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China. Electronic address: gaoweiping@tsinghua.edu.cn.
authors.forename: Weiping
authors.initials: W
authors.lastname: Gao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D003545:Cysteine / Q000737:chemistry; D003553:Cystine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D019085:Fluorescent Antibody Technique, Direct; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry*; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D041681:NIH 3T3 Cells; D011092:Polyethylene Glycols / Q000737:chemistry*; D011487:Protein Conformation; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis / Q000276:immunology; D012235:Rhodamines / Q000032:analysis* / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D000068878:Trastuzumab / Q000737:chemistry / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D003545:Cysteine; D003553:Cystine; D004220:Disulfides; D005456:Fluorescent Dyes; D018796:Immunoconjugates; D008689:Methacrylates; D011092:Polyethylene Glycols; D018719:Erb-b2 Receptor Tyrosine Kinases; D012235:Rhodamines; D000068878:Trastuzumab; C524499:polyethylene glycol methacrylate; C508053:ERBB2 protein, human; C029773:rhodamine B
keywords: Antibody conjugate; Antigen detection; Polymer conjugate; Signal amplification