Functional expression of single-chain antibody to leukotriene C(4).
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).
Kawakami
Yuki
Y
Department of Nutritional Science, Okayama Prefectural University, Okayama 719-1197, Japan.
Yamashita
Chiaki
C
Kashiwase
Yoshiko
Y
Morinaka
Tomoko
T
Suzuki-Yamamoto
Toshiko
T
Yamashita
Hiromi
H
Kimoto
Masumi
M
Tsuji
Hideaki
H
Kurahashi
Yuko
Y
Daiyasu
Hiromi
H
Toh
Hiroyuki
H
Sugahara
Mitsuaki
M
Miyano
Masashi
M
Yamamoto
Shozo
S
Takahashi
Yoshitaka
Y
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
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D017997:Leukotriene C4