title: Identification of Immunoglobulin Gene Sequences from a Small Read Number of mRNA-Seq Using Hybridomas.
abstract: Identification of immunoglobulin genes in hybridomas is essential for producing antibodies for research and clinical applications. A couple of methods such as RACE and degenerative PCR have been developed for determination of the Igh and Igl/Igk coding sequences (CDSs) but it has been difficult to process a number of hybridomas both with accuracy and rapidness. Here, we propose a new strategy for antibody sequence determination by mRNA-seq of hybridomas. We demonstrated that hybridomas highly expressed the Igh and Igl/Igk genes and that de novo transcriptome assembly using mRNA-seq data enabled identification of the CDS of both Igh and Igl/Igk accurately. Furthermore, we estimated that only 30,000 sequenced reads are required to identify immunoglobulin sequences from four different hybridoma clones. Thus, our approach would facilitate determining variable CDSs drastically.
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.|Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
authors: Yuki
authors: Y
authors: Kuniyoshi
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
authors: Kazumitsu
authors: K
authors: Maehara
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
authors: Takeshi
authors: T
authors: Iwasaki
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
authors: Masayasu
authors: M
authors: Hayashi
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
authors: Yuichiro
authors: Y
authors: Semba
authors: Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
authors: Masatoshi
authors: M
authors: Fujita
authors: Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors: Yuko
authors: Y
authors: Sato
authors: Department of Biological Sciences, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
authors: Hiroshi
authors: H
authors: Kimura
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
authors: Akihito
authors: A
authors: Harada
authors: Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
authors: Yasuyuki
authors: http://orcid.org/0000-0001-6440-9954
authors: Y
authors: Ohkawa
mesh_terms: D020869:Gene Expression Profiling; D005803:Genes, Immunoglobulin / Q000235:genetics*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D017423:Sequence Analysis, RNA
chemical_list: D012333:RNA, Messenger