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Determination of ß-Lactoglobulin in Complex Systems by a Simple Immunological Procedure¹

Laboratory of Biochemistry, Department of Dairy Science, University of Illinois, Urbana

ABSTRACT

A simple method, based on immunological procedures described originally by Oudin, has been developed to quantitate the amount of ß-lactoglobulin present in complex systems. The solution containing ß-lactoglobulin is allowed to diffuse into an agar layer containing antisera to the ß-lactoglobulin. The distance of this migration, as measured by the formation of a turbid zone in the tube, is proportional to the ß-lactoglobulin concentration. The usable range of the method is about 10–80 µg/ml. Since milk contains about 2,500 µg/ml of ß-lactoglobulin, dilution is the only sample preparation required for milk and liquid milk products. The ß-lactoglobulin in solid materials must first be solubilized in a suitable tissue homogenizer.

A concentration procedure is also described with which it has been possible to determine less than 1 µg/ml of ß-lactoglobulin in a complex nutrient medium used to maintain bovine mammary secretory cells in laboratory culture and containing levels of other proteins up to ten-thousand times more concentrated than the ß-lactoglobulin.

¹ This investigation was supported by a grant from the National Science Foundation.