Hen egg yolk antibodies purified by antigen affinity under highly alkaline conditions provide new tools for diagnostics. Human intact parathyrin as a model antigen.

Abstract

Hen egg yolks have been recognized as convenient source for specific antibodies, although their utility in diagnostic applications has been hampered by the lack of efficient purification methods. In the present study, anti-human parathyrin antibodies were raised in rabbits, hens and mice using synthetic human parathyrin peptide (1-84) as an antigen. The antibodies were affinity purified with amino- (1-30) and carboxy- (49-89) terminal peptides of parathyrin under highly alkaline elution conditions, and were evaluated for their diagnostic value in different combinations in immunoenzymometric assay (IEMA) format. A pair of hen egg yolk antibodies were subjected to further methodological validation in the IEMA that was constructed with the N-terminal capture and C-terminal detection antibodies. The synthetic intact human parathyrin (1-84) peptide served as a standard. This within-day IEMA procedure turned out to be sensitive and it correlated well with the two independent intact parathyrin immunoradiometric assays. As shown in the present study, the immuno affinity purification with the highly alkaline elution conditions provides an efficient method for utilization of hen egg yolk antibodies. This is the first report on an application making use of a combination of two hen egg yolk antibody preparations in measuring a homogeneous protein in human serum.