Abstract
Diagnosis of many infectious, autoimmune diseases and cancers depends on the detection of specific antibodies against peptide epitope by enzyme-linked immunosorbent assay (ELISA). However, small peptides are difficult to be coated on the plate surfaces. In this study, we selected GnRH as a model hapten to evaluate whether VEGF121 would be suitable as an irrelevant hapten-carrier to develop a universal platform for specific antibodies detection. Firstly, GnRH was fused to the C terminus of VEGF121 and the resultant fusion protein VEGF–GnRH expressed effectively as inclusion bodies in Escherichia coli. Thereafter, VEGF–GnRH was easily purified to near homogeneity with a yield of about 235 mg from 2.1 L induced culture. At last, VEGF–GnRH was used to perform ELISA and western blot, and our results suggested that VEGF–GnRH was capable of detecting anti-GnRH antibodies in sera both qualitatively and quantitatively. Indeed, previous studies of our laboratory had demonstrated that other fusion proteins such as VEGF–Aβ10, VEGF–GRP, VEGF–CETPC, and VEGF–βhCGCTP37 were able to detect their corresponding antibodies specifically. Therefore, VEGF121 may be a suitable irrelevant fusion partner of important diagnostic peptide markers. Our works would shed some light on the development of a universal platform for detection of specific antibodies.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 81101706) and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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The authors declare that they have no conflict of interest.
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Wang, X.J., Zhou, L.J., Zhu, X.J. et al. Detection of antibodies against customized epitope: use of a coating antigen employing VEGF as fusion partner. Appl Microbiol Biotechnol 98, 6659–6666 (2014). https://doi.org/10.1007/s00253-014-5618-6
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DOI: https://doi.org/10.1007/s00253-014-5618-6