Monoclonal Antibody Production and the Development of an Indirect Competitive Enzyme-Linked Immunosorbent Assay for Screening Spiramycin in Milk

Abstract

To monitor spiramycin (SP) residue in milk, a monoclonal antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed. This study described the preparation of three immunogens and the production of a high-affinity mAb. After optimization, the 50% inhibition concentration (IC50) for the developed icELISA was estimated as 0.97 ng/mL in the assay buffer, and the limit of detection and limit of quantitation were 2.51 and 4.40 μg/L in the milk matrix. The newly developed assay demonstrated negligible cross-reactivity with 15 other macrolide antibiotics, but not with kitasamycin (23.4%). The mean recoveries ranged from 81 to 103% for the spiked samples (5, 10, and 50 μg/L), and the coefficient of variation ranged from 5.4 to 9.6%. The icELISA was validated by LC-MS/MS method, and all results demonstrated that it was a suitable screening method for detecting SP residue in milk without requiring a cleanup process.

A novel immunoassay for quantitative drug abuse screening in serum

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Gold immunochromatographic assay for kitasamycin and josamycin residues screening in milk and egg samples

“…Hapten and antigen synthesis The hapten was synthesized using the CMO method (Figure 2) (Jiang et al, 2013). Onehundred mg of KIT and 20 mg of CMO were added to a mixture of 5 mL methanol and 5 mL 0.02 M sodium bicarbonate solution.…”

Botryoid-shaped nanoparticles-enhanced ELISA for ochratoxin A

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