Enzyme-linked immunosorbent assay development for advanced glycation end products and brochocin-C

Advanced glycation end products (AGEs) are important components to detect in diabetic patients. Polyclonal antiserum against both physiological and non-physiological AGEs was generated. Following a competitive enzyme-linked immunosorbent assay (ELISA) with the antiserum, the absorbance maximum could be reduced 50% using 89.6 ppm glucose glycated ovalbumin. The antiserum seemed to recognize AGE crosslinks that were cleaved by N-phenacylthiazolium bromide, however, simple glycated proteins were not recognized.Attempts to develop an ELISA procedure for brochocin-C, an anti-botulinal bacteriocin, were not successful. Large losses of brochocin-C using standard purification procedures were traced to butanol extraction and subsequent size exclusion chromatography due to extensive agglomeration of brochocin-C. Polyclonal antiserum developed against brochocin-C conjugate failed to provide useful antibodies for ELISA development due to the lack of sufficient purified brochocin-C. Polyclonal antiserum against carboxy-terminal synthetic peptide of brochocin-C weakly recognized the brochocin-C conjugate, but polyclonal antiserum against amino-terminal peptide did not.