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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 34, Number 2
BKCSDE 34(2)
February 20, 2013 

 
Title
Development of a Single Chain Antibody Using a Phage Display Cloning Method for the Detection of 2,4-Dinitrotoluene
Author
Jung-Hyun Na, Man-Seok Joo, Won-Kyu Lee, Hyunbo Shim, Si-Hyung Lim, Sang Taek Jung, Yeon Gyu Yu*
Keywords
DNT, Sensor, Single chain antibody, Biopanning, Phage display
Abstract
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.
Page
460 - 464
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