Bulletin of the Korean chemical society | |
Development of a Single Chain Antibody Using a Phage Display Cloning Method for the Detection of 2,4-Dinitrotoluene | |
Si-Hyung Lim1  Hyunbo Shim1  Sang Taek Jung1  Won-Kyu Lee1  Jung-Hyun Na1  Yeon Gyu Yu1  Man-Seok Joo1  | |
关键词: DNT; Sensor; Single chain antibody; Biopanning; Phage display; | |
DOI : | |
学科分类:化学(综合) | |
来源: Korean Chemical Society | |
【 摘 要 】
Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with KD value of 6.0 × 10−7 M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.
【 授权许可】
Unknown
【 预 览 】
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RO201912010244346ZK.pdf | 3344KB | download |