Standardisation of Pollen Extracts by Gel Diffusion

Quantitative Gel Diffusion Methods for Assay of Antigens and Antibodies

“…) (2) Direct assay by the reversed gradient diffusion method: Ag-gradient method. This test has been developed similarly to the gradient dif fusion method for measuring antigens (Augustin and Hayward, 1955), but, instead of an Ab-gradient across the middle layer, an Aggradient is established by incorporating Ag in the bottom gel layer, instead of Ab; Ab instead of Ag constitutes the top layer.…”

“…The height of this neutral column (containing neither reactant) is carefully adjusted by holding each tube against a reference tube during the filling process. A measuring aid may be used (Augustin and Hayward, 1955) but is not required after a little practice. In this way, many tubes may be rapidly filled, all with columns of equal height.…”

“…1 shows that this method makes it possible to maintain high Abtitres for at least one year, with only three sessions of multiple in jections on one day every three months, and frequent bleedings. Single assays were carried out on each bleeding by the optimal proportion method of Dean and Webb (1926) and by the gradient double diffusion method (Augustin and Hayward, 1955). Sera containing 0.02% Na-azidc as a preservative were either stored at 4° C, or freeze dried for storage at 4° C. There was no deterioration on freeze drying and the titres of the two preparations with respect to a standard Ag-solution and in comparison with each other remained constant throughout the vear.…”

Fundamental Aspects of Single Versus Double Diffusion Methods for Immunological Assays

“…The most reliable of these methods is probably that by Feinberg, who has gone to great trouble to stan dardise this procedure by carefully choosing his Petri dishes and employing a special cutter manufactured with great accuracy for cutting out the Ag and Ab reservoirs. The principle of the test has been taken from our gradient double diffusion method (Augustin and Hayward, 1955), i.e. an Ab gradient is established from a central Ab cup before filling the exactly equidistant peripheral cups with Ag solutions; once more stationary lines are formed.…”

The Nature and Properties of Antibodies in Sera of Allergic Individuals. A Review

“…The different attempts to use the various forms of anaphylaxis, such as passive sensitization of animals with allergic antibodies (26,27), the S c h u l t z -D a le technique (28) and passive cutaneous anaphylaxis (29,30) for the detection of allergic antibodies have generally met with failure, although a few positive reactions have been reported (29). The in vitro combination of antibodies of allergic sera with the correspond ing allergens has been demonstrated by complement fixation studies (31,32), more consistently with sera from treated allergic individ uals.…”