268
Views
17
CrossRef citations to date
0
Altmetric
Original Articles

Production of antibodies and development of enzyme-linked immunosorbent assay for valnemulin in porcine liver

, , , , &
Pages 244-252 | Received 04 Aug 2012, Accepted 30 Sep 2012, Published online: 01 Nov 2012
 

Abstract

Polyclonal and monoclonal antibodies against valnemulin, a new semi-synthetic antibiotic derivative of pleuromutilin administered to treat swine dysentery and pneumonia, were generated. To achieve high enzyme-linked immunosorbent assay sensitivity for valnemulin, several heterologous coating antigens were prepared and evaluated, differing in the length of the spacer arm and the conjugation site between valnemulin and carrier protein. After the optimisation of immuno-reagents dilution, the enzyme-linked immunosorbent assay, based on polyclonal antibody number 2 and one heterologous coating antigen, showed the highest sensitivity with an IC50 value for valnemulin of 0.96 ng/mL in buffer. For spiked porcine liver, an extraction procedure with a mixture of acetonitrile and 0.01 M hydrochloric acid (40:60 v/v) was proposed and no further sample pre-treatment other than 10 times dilution of the extract was necessary prior to analysis, which gave recovery values ranging from 75.7% to 89.4%. The dynamic assay range and the limit of detection of the assay were 2.4−49.9 and 1.67 µg/kg for porcine liver, respectively. The assay was compared with a confirmation method based on LC-MS/MS by using valnemulin-treated samples, and a satisfactory correlation between both methods was observed.

Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (no. 30901086), State Key Program of National Natural Science of China (no. 30830082) and key projects in the National Science & Technology Program during the twelfth Five-Year Plan Period (no. 2011BAK10B01).

Log in via your institution

Log in to Taylor & Francis Online

PDF download + Online access

  • 48 hours access to article PDF & online version
  • Article PDF can be downloaded
  • Article PDF can be printed
USD 61.00 Add to cart

Issue Purchase

  • 30 days online access to complete issue
  • Article PDFs can be downloaded
  • Article PDFs can be printed
USD 799.00 Add to cart

* Local tax will be added as applicable

Related Research

People also read lists articles that other readers of this article have read.

Recommended articles lists articles that we recommend and is powered by our AI driven recommendation engine.

Cited by lists all citing articles based on Crossref citations.
Articles with the Crossref icon will open in a new tab.