A method for estimation of the relative antigenic potencies of preparations containing common new antigens derived from a precursor protein (β-lactoglobulin)

Chemistry of Allergens

“…Table I contains a summary of the yields, nitrogen contents and per centage of the total nitrogen in each of the dialysate fractions (D1-D6) obtained in the successive hydrolyses of /J-lactoglobulin [16]. D2 and D3 were used for the isolation of the purified new antigens because they not only contained the new antigens common to all of the dialysates [16,17] but they also were available in greater quantity than the other fractions. Figure 1A, B shows the desalting and partial fractionation of D2 and D3 on Bio-Gel P-2 into three fractions, A, B, and C, respectively.…”

“…None of the antigens reacted nonspecifically with strips from nonsensitized guinea pigs in the quantities used in the challenge doses. To insure that re actions on sensitized tissues were caused by new antigens and not by /J-lactoglobulin and/or an autodigest of pepsin, all tests for new antigen were made on strips that either did not react to or were first desensitized to a mixture of /?-lactoglobulin and the autodigest of pepsin (PEPD) [15][16][17]. Almost all strips from animals sensitized with the purified polypeptides did not react to /Mactoglobulin and PEPD.…”

“…The basic Schultz-Dale technique used has been de scribed by Coulson [3] and in our previous applications of the method [15][16][17]. Virgin female guinea pigs, weighing about 225 g, were sensitized on the same day by subcutaneous injections (nuchal area) with two 0.5-ml volumes of antigen emulsified with Freund's complete adjuvant.…”