title: Development of a novel semi-quantitative analysis system for ultramicroscale samples by fluorescent capillary isoelectric focusing.
abstract: Microchemistry provides methods to analyze small quantities of chemical substances, including proteins, nucleic acids, and carbohydrates in various fields such as biomedical research, tissue engineering, molecular biology, and regeneration medicine. We therefore developed a fluorescent capillary isoelectric focusing (fluorescent cIEF) system for protein detection at an ultramicroscale volume, which aimed to isolate and identify, from a heterogeneous mixture of transduced cells, induced pluripotent stem cells (iPSCs) that could be fully reprogrammed. In addition, we demonstrated that the SOX2 protein, which is indispensable for the acquisition of pluripotency, could be detected by this new fluorescent cIEF system to identify iPSCs in the early phase of complete reprogramming. This method took less than 1 h for completion, including the time required for the antibody-antigen (Ab-Ag) reaction, and required as few as approximately three cells. Thus, this system could help improve iPSC generation as well as cut costs and reduce workloads.
authors: Department of Regenerative Medicine, Kyoto Prefectural University of Medicine, Japan.
authors: Daisuke
authors: D
authors: Kami
authors: Faculty of Engineering, Yokohama National University, Japan.
authors: Makoto
authors: M
authors: Takahashi
authors: Department of Regenerative Medicine, Kyoto Prefectural University of Medicine, Japan. Electronic address: gojos@koto.kpu-m.ac.jp.
authors: Satoshi
authors: S
authors: Gojo
authors: Department of Vascular Medicine, Tokyo Metropolitan Institute of Gerontology, Japan.
authors: Masashi
authors: M
authors: Toyoda
authors: Metaboscreen Co., Ltd., Japan.
authors: Ryoko
authors: R
authors: Aso
authors: Metaboscreen Co., Ltd., Japan.
authors: Ryuichi
authors: R
authors: Sekizawa
authors: Faculty of Engineering, Yokohama National University, Japan.
authors: Masatoshi
authors: M
authors: Watanabe
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D002417:Cattle; D002469:Cell Separation / Q000295:instrumentation; D004867:Equipment Design; D005453:Fluorescence; D006367:HeLa Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D007525:Isoelectric Focusing / Q000295:instrumentation*; D008832:Microchemistry / Q000295:instrumentation; D055748:SOXB1 Transcription Factors / Q000032:analysis; D012710:Serum Albumin, Bovine / Q000032:analysis
chemical_list: C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D012710:Serum Albumin, Bovine
keywords: Capillary isoelectric focusing; High sensitive analysis; Induced pluripotent stem cell; Microchemistry