Specific antibody for pesticide residue determination produced by antibody-pesticide complex

Abstract

A new method for specific antibody production was developed using antibody (Ab)-pesticide complex as a unique immunogen. Parathion (PA) was the targeted pesticide, and rabbit polyclonal antibody (Pab) and mouse monoclonal antibody (Mab) were used as carrier proteins. The Ab-PA complexes were generated by conjugating the two antibodies with an excessive dosage of PA. It was shown that the sensitivity of homologous enzyme-linked immunosorbent assay (ELISA) using the new antibodies was similar to that using original antibodies. However, the new mouse Pab had not only similar positive recognizing spectrum as the original Mab, but also a significantly improved sensitivity in heterologous ELISA when some recognizable competitors were applied. IC₅₀ value of ELISA based on a combination of the new mouse Pab and hapten 9 was 0.24 ng/mL, which was 445.54 times as that of the homologous ELISA. An Ab-pesticide complex may be a suitable alternative immunogen for producing highly specific antibody to improve the immunoassay sensitivity.

Keywords:

• Antibody-pesticide complex
• parathion
• ELISA
• assay sensitivity

Acknowledgments

This work was supported by the National Natural Science Foundation of China (NSFC No. 30671391) and the Key Projects in the National Science & Technology Pillar Program during the Eleventh Five-year Plan Period (NO2009BADB9B09).

Notes

^a •••: conjugates showed high reactivity (A ≥ 1.5) with the antibody; ••○: conjugates showed middle reactivity (1.0 ≤ A < 1.5) with the antibody; •○○: conjugates showed low reactivity (0.5 ≤ A < 1.0) with the antibody; ○○○: conjugates showed no reactivity (A < 0.5) with the antibody.

^b:No ELISA curve could be obtained.