Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1933003 | Biochemical and Biophysical Research Communications | 2010 | 5 Pages |
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.