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J. Biochem, 1983, Vol. 94, No. 1 43-49
© 1983 Japanese Biochemical Society

research-article

Blocking Effect of Trypsin Associated with Ovomucoid on the Antibody Binding to Ovomucoid

Tsukasa MATSUDA, Kenji WATANABE and Ryo NAKAMURA

Department of Food Science and Technology, Faculty of Agriculture, Nagoya University Chikusa-ku, Nagoya, Aichi 464

Ovomucoid-trypsin association complex was prepared by incubating chicken egg white ovomucoid with bovine trypsin. The reactivity of ovomucoid-trypsin complex was investigated by immunodiffusion, quantitative precipitation and enzyme-linked immunosorbent assay (ELISA). It was demonstrated that the association of trypsin with ovomucoid hindered the binding of the specific antibody at some antigenic sites of ovomucoid by lowering the antibody-binding affinity of these sites. The anti-ovomucoid antiserum was absorbed with ovomucoid-trypsin complex, and non-absorbed antibody was collected by immunoaffinity chromatography of ovomu-coid-coupled Sepharose 4B. The antibody blocked the trypsin-inhibitory activity of ovomucoid in a molar ratio (antibody/ovomucoid) of about 1.2 : 1. The findings suggested that at least one antigenic site is located near the reactive site of trypsin inhibition Formula of ovomucoid.


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