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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 22, Number 9
BKCSDE 22(9)
September 20, 2001 

Influence of Coating Ligands on Enzyme-linked Immunosorbent Assay of Toluene
So Young Kim, Nam Taek Lee, Myung Ja Choi
Coating ligand, Competitive ELISA, Toluene, Benzene, Xylene.
The specificity of conjugation site for coating ligands was investigated using toluene-bovine serum albumin (BSA) conjugates in which BSA was conjugated at the position of o-, m-, and ρ-toluic acid. Toluene-BSA conjugated at ρ-position showed a binding activity of about 89-95%, whereas, those conjugated at o- and m-position of toluene exhibited a binding activity of 5 and 11%, respectively. On the basis of the above result, coating ligands with various proteins (OVA, BSA, KLH) were compared by conjugating with ρ-toluic acid, and toluene-KLH was considered as the best coating ligand for this ELISA. Indirect competitive ELISA method was developed using anti-toluene antibody and ρ-position conjugated toluene-KLH. The dose-response curve constructed after kinetic and optimization studies showed a 1×10-4 - 1×102 mM detectable response range with 0.1 μM detectability. In specificity test of the antibody, the binding capabilities of aromatic compounds substituted with nitro-, alkyl-, chloro-, and hydroxyl group were larger rather than those of aromatic compounds (benzene, toluene and xylene) themselves. Also, tests with soil and water samples that had been spiked with toluene resulted in 102.7-113.7% recovery.
953 - 957
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