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Development of a single chain antibody using a phage display cloning method for the detection of 2,4-dinitrotoluene

Title
Development of a single chain antibody using a phage display cloning method for the detection of 2,4-dinitrotoluene
Authors
Na J.-H.Joo M.-S.Lee W.-K.Shim H.Lim S.-H.Taek Jung S.Gyu Yu Y.
Ewha Authors
심현보
SCOPUS Author ID
심현보scopus
Issue Date
2013
Journal Title
Bulletin of the Korean Chemical Society
ISSN
0253-2964JCR Link
Citation
vol. 34, no. 2, pp. 460 - 464
Indexed
SCI; SCIE; SCOPUS; KCI WOS scopus
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.
DOI
10.5012/bkcs.2013.34.2.460
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일반대학원 > 바이오융합과학과 > Journal papers
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