title: In situ drug-receptor binding kinetics in single cells: a quantitative label-free study of anti-tumor drug resistance.
abstract: Many drugs are effective in the early stage of treatment, but patients develop drug resistance after a certain period of treatment, causing failure of the therapy. An important example is Herceptin, a popular monoclonal antibody drug for breast cancer by specifically targeting human epidermal growth factor receptor 2 (Her2). Here we demonstrate a quantitative binding kinetics analysis of drug-target interactions to investigate the molecular scale origin of drug resistance. Using a surface plasmon resonance imaging, we measured the in situ Herceptin-Her2 binding kinetics in single intact cancer cells for the first time, and observed significantly weakened Herceptin-Her2 interactions in Herceptin-resistant cells, compared to those in Herceptin-sensitive cells. We further showed that the steric hindrance of Mucin-4, a membrane protein, was responsible for the altered drug-receptor binding. This effect of a third molecule on drug-receptor interactions cannot be studied using traditional purified protein methods, demonstrating the importance of the present intact cell-based binding kinetics analysis.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: 1] Center for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA [2] School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044, China.
authors.forename: Linliang
authors.initials: L
authors.lastname: Yin
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Gonzalez-Malerva
authors.affiliation: Center for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Shaopeng
authors.initials: S
authors.lastname: Wang
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Yu
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Seron
authors.initials: S
authors.lastname: Eaton
authors.affiliation: School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 400044, China.
authors.forename: Shengtao
authors.initials: S
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
authors.forename: Hong-Yuan
authors.initials: HY
authors.lastname: Chen
authors.affiliation: Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
authors.affiliation: 1] State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China [2] Center for Bioelectronics and Biosensors, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA [3] Department of Electrical Engineering, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Nongjian
authors.initials: N
authors.lastname: Tao
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D004347:Drug Interactions; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005260:Female; D006801:Humans; D055264:Mucin-4 / Q000378:metabolism; D011485:Protein Binding; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D055264:Mucin-4; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C508053:ERBB2 protein, human