title: Functional expression of single-chain antibody to leukotriene C(4).
abstract: Leukotriene C(4) (LTC(4)) is synthesized by binding of glutathione to LTA(4), an epoxide derived from arachidonic acid, and further metabolized to LTD(4) and LTE(4). We previously prepared a monoclonal antibody with a high affinity and specificity to LTC(4). To explore the structure of the antigen-binding site of a monoclonal antibody against LTC(4) (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 LTC(4) comparable to mAbLTC. The scFvLTC also bound to LTD(4) and LTE(4) with 48% and 17% reactivities, respectively, as compared with LTC(4) binding, whereas the antibody showed almost no affinity for LTB(4).
authors.affiliation: Department of Nutritional Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kawakami
authors.forename: Chiaki
authors.initials: C
authors.lastname: Yamashita
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Kashiwase
authors.forename: Tomoko
authors.initials: T
authors.lastname: Morinaka
authors.forename: Toshiko
authors.initials: T
authors.lastname: Suzuki-Yamamoto
authors.forename: Hiromi
authors.initials: H
authors.lastname: Yamashita
authors.forename: Masumi
authors.initials: M
authors.lastname: Kimoto
authors.forename: Hideaki
authors.initials: H
authors.lastname: Tsuji
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kurahashi
authors.forename: Hiromi
authors.initials: H
authors.lastname: Daiyasu
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Toh
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Sugahara
authors.forename: Masashi
authors.initials: M
authors.lastname: Miyano
authors.forename: Shozo
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Takahashi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis* / Q000235:genetics; D017997:Leukotriene C4 / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D017997:Leukotriene C4