title: Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes.
abstract: Exosome-mediated signal transportation plays a variety of critical roles in cancer progression and metastasis. From the aspect of cancer diagnosis, circulating exosomes are ideal resources of biomarkers because molecular features of tumor cells are transcribed on them. However, isolating pure exosomes from body fluids is time-consuming and still major challenge to be addressed for comprehensive profiling of exosomal proteins and miRNAs. Here we constructed anti-CD9 antibody-coupled highly porous monolithic silica microtips which allowed automated rapid and reproducible exosome extraction from multiple clinical samples. We applied these tips to explore lung cancer biomarker proteins on exosomes by analyzing 46 serum samples. The mass spectrometric quantification of 1,369 exosomal proteins identified CD91 as a lung adenocarcinoma specific antigen on exosomes, which was further validated with CD9-CD91 exosome sandwich ELISA measuring 212 samples. Our simple device can promote not only biomarker discovery studies but also wide range of omics researches about exosomes.
authors.affiliation: 1] Division of Biosciences, Functional Proteomics Center, Graduate School of Frontier Sciences, the University of Tokyo, Tokyo, Japan [2] Laboratory for Genome Sequencing Analysis, Center for Integrated Medical Sciences, RIKEN, Tokyo, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Ueda
authors.affiliation: Department of Molecular and Internal Medicine, Hiroshima University, Hiroshima, Japan.
authors.forename: Nobuhisa
authors.initials: N
authors.lastname: Ishikawa
authors.affiliation: Laboratory for Genome Sequencing Analysis, Center for Integrated Medical Sciences, RIKEN, Tokyo, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Tatsuguchi
authors.affiliation: 1] Division of Biosciences, Functional Proteomics Center, Graduate School of Frontier Sciences, the University of Tokyo, Tokyo, Japan [2] Laboratory for Genome Sequencing Analysis, Center for Integrated Medical Sciences, RIKEN, Tokyo, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Saichi
authors.affiliation: 1] Division of Biosciences, Functional Proteomics Center, Graduate School of Frontier Sciences, the University of Tokyo, Tokyo, Japan [2] Laboratory for Genome Sequencing Analysis, Center for Integrated Medical Sciences, RIKEN, Tokyo, Japan.
authors.forename: Risa
authors.initials: R
authors.lastname: Fujii
authors.affiliation: Laboratory for Genome Sequencing Analysis, Center for Integrated Medical Sciences, RIKEN, Tokyo, Japan.
authors.forename: Hidewaki
authors.initials: H
authors.lastname: Nakagawa
mesh_terms: D000230:Adenocarcinoma / Q000097:blood* / Q000175:diagnosis; D000328:Adult; D055910:Antibodies, Immobilized / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D055354:Exosomes / Q000378:metabolism*; D005260:Female; D006801:Humans; D007118:Immunoassay; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D008175:Lung Neoplasms / Q000097:blood* / Q000175:diagnosis; D008297:Male; D012372:ROC Curve; D012822:Silicon Dioxide / Q000737:chemistry*; D053719:Tandem Mass Spectrometry
chemical_list: D055910:Antibodies, Immobilized; D014408:Biomarkers, Tumor; C475476:LRP1 protein, human; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D012822:Silicon Dioxide