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Functional expression of single-chain antibody to leukotriene C4 |
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| Authors: | Yuki Kawakami Yoshiko Kashiwase Toshiko Suzuki-Yamamoto Masumi Kimoto Yuko Kurahashi Hiroyuki Toh Masashi Miyano Yoshitaka Takahashi |
| Institution: | a Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, 111 Kuboki, Soja, Okayama 719-1197, Japan b Department of Food Science and Nutrition, Faculty of Human Life and Science, Doshisha Women’s College of Liberal Arts, Kamigyou-ku, Kyoto 602-0893, Japan c The Center for Advanced Medical Engineering and Informatics, Osaka University, Suita, Osaka 565-0871, Japan d Division of Bioinformatics, Medical Institute of Bioregulation, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan e Structural Biophysics Laboratory, RIKEN SPring-8 Center, Harima Institute, Sayo, Hyogo 679-5148, Japan |
| Abstract: | Leukotriene C4 (LTC4) is synthesized by binding of glutathione to LTA4, an epoxide derived from arachidonic acid, and further metabolized to LTD4 and LTE4. We previously prepared a monoclonal antibody with a high affinity and specificity to LTC4. To explore the structure of the antigen-binding site of a monoclonal antibody against LTC4 (mAbLTC), we isolated full-length cDNAs for heavy and light chains of mAbLTC. The heavy and light chains consisted of 461 and 238 amino acids including a signal peptide with molecular weights of 51,089 and 26,340, respectively. An expression plasmid encoding a single-chain antibody comprising variable regions of mAbLTC heavy and light chains (scFvLTC) was constructed and expressed in COS-7 cells. The recombinant scFvLTC showed a high affinity with LTC4 comparable to mAbLTC. The scFvLTC also bound to LTD4 and LTE4 with 48% and 17% reactivities, respectively, as compared with LTC4 binding, whereas the antibody showed almost no affinity for LTB4. |
| Keywords: | Single-chain antibody Leukotriene C4 Arachidonic acid Monoclonal antibody |
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