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Development of a highly sensitive icELISA to detect semicarbazide based on a monoclonal antibody

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A highly sensitive and specific monoclonal antibody was prepared to detect semicarbazide (SEM). Hapten 4-{[(aminocarbonyl)hydrazono]methyl}benzoic acid (4-CPSEM) was synthesised through the condensation reactions of SEM and 4-carboxybenzaldehyde (4-CBA). The active ester method was employed to couple the hapten to bovine serum albumin, keyhole limpet hemocyanin and ovalbumin (OVA) to obtain conjugates of immunogens and coating antigens. A novel hapten 2-[(aminocarbonyl)hydrazono]acetic acid (SEM-A) was also synthesised from SEM and oxoacetic acid to prepare a heterologous coating antigen (SEM-A-OVA). The linear ranges of inhibition curves in the optimised indirect competitive enzyme-linked immunosorbent assay (icELISA) method were in the range of 0.0071–0.056 ng mL−1 and 0.018–0.209 ng mL−1 for coating antigens SEM-A-OVA and 4-CPSEM-OVA, respectively. The IC50 was 0.019 ng mL−1 for SEM (SEM in the form of 4-nitrobenzaldehyde semicarbazone) and 0.13 ng mL−1 for 2-nitrobenzaldehyde semicarbazone, both of which were well below the minimum required performance limit of 1 µg·kg−1 set by the European Commission.
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Keywords: indirect competitive enzyme-linked immunosorbent assay; monoclonal antibody; nitrofurazone; semicarbazide

Document Type: Research Article

Affiliations: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, PR China

Publication date: May 4, 2015

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