Preparation of monoclonal antibody against bovine .BETA.-lactoglobulin and its unique binding affinity.
- 1 January 1987
- journal article
- Published by Oxford University Press (OUP) in Agricultural and Biological Chemistry
- Vol. 51 (3) , 797-802
- https://doi.org/10.1271/bbb1961.51.797
Abstract
Five hybridoma cell lines, 21B3, 31A4, 61C1, 61B4, and 62A6 were established and characterized to produce IgG1 antibodies with specificity for bovine β-1actoglobulin (β-LG) which is a potent milk allergen. A representative monoclonal antibody (MAb) 21B3 had a unique affinity; it bound more strongly to RCM (reduced carboxymethylated) β-LG than to native β-LG. The affinity increased drastically when β-LG was heated at 70°C or above for 10min at pH 7.0, while β-LG heated at pH 3.0 did not alter its affinity even when heated at 80°C for 10min. An enzymeprobe method suggested that unfolding of β-LG by heating at pH 7.0 seemed to start at 70°C, and β-LG heated at 90°C and RCM β-LG were extensively unfolded. The results indicated that the phase of unfolding and increase of affinity by heating were coincidental. Finally the relationship of the unique affinity of MAb and the β-LG conformation was discussed.Keywords
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