Journal
FOOD AND AGRICULTURAL IMMUNOLOGY
Volume 26, Issue 3, Pages 356-365Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/09540105.2014.914891
Keywords
semicarbazide; nitrofurazone; monoclonal antibody; indirect competitive enzyme-linked immunosorbent assay
Funding
- Key Programs from MOST [2012AA06A303, 2012BAD29B04]
- Jiangsu Province
- MOF
- MOE [201310135, JUSRP51308A]
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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 mu g center dot kg(-1) set by the European Commission.
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