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Synthesis and identification of artificial mercury chelate antigen |
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Abstract In this paper, artificial hapten Hg-ITCBE was synthesized using ITCBE (Isothiocyanobenzylethyl enediamine tetraacetic acid) as the bifunctional chelating agent. Antigens were synthesized through the coupling the hapten with carrier protein keyhole limpet hemocyanin(KLH) and bovine serum albumin (BSA). After the assay for concentration of the antigen with bicinchoninic acid (BCA) method, the hapten antigen and carrier protein were scanned with ultraviolet spectrophotometer, and verified with SDS-PAGE. The degree of conjugate substitution was quantified using the trintrobenzenesulfonic acid(TNBS) method. Concentration of mercury ion in antigen determined with graphite-furnace atomic absorption spectrometry(AAS). The results indicated antigens were synthesized successfully. The extent of substitution of free lysine residues was 34.75 ± 4.60% for Hg-ITCBE-KLH, 40.61 ±0.99%for the Hg-ITCBE-BSA and 61.27 ±0.69% for the ITCBE-BSA. The concentration of mercury ion were 38.4±0.5μg/ml, 125.5±0.9μg/ml and 0μg/ml , respectively.
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Received: 30 September 2007
Published: 25 February 2008
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