Separation of antigens by immunological specificity. 1. Method for separating individual antigen-antibody complexes from mixed antigens and antibodies

Immunogenicity and molecular characterisation of staphylococcal haemolysin (Plate XIII)

“…After electrophoresis, troughs were filled with sheep anti-rabbit antiserum (Stewart-Tull and Rowe, 1975) or 6 haemolysin (200 pg/ml) and lines of precipitation were allowed to develop for 24 h. Immunodiffusion was by the method of Ouchterlony (1968) with wells of 1 1-mm diameter spaced 1 1 mm between centres or, by a micromethod, with wells of 3-mm diameter spaced 6 mm between centres. Quantitative precipitin analysis on antisera was done as described by Smith et al (1962) and the protein content of precipitates was measured by the method of Lowry et al (1951). The diffusion coefficient of 6 haemolysin was estimated as described by Allison and Humphrey (1960) and the mean of three measurements was taken.…”

Separation of antigens by immunological specificity. Use of cellulose-linked antibodies as immunosorbents

“…It should be emphasized that the method does not require the pre-existence of pure antigens to obtain monospecific antisera (Smith et al 1962(Smith et al , 1964 …”

“…Antigens Fluorescent ovalbumin. This was prepared as described by Smith et al (1962), and further purified. An 18ml.…”

“…of 0.9% NaCl) on days 41, 43 and 45. Rabbits were bled on day 50 and y-globulin concentrates were prepared as described by Smith et al (1962). Approx.…”

“…Initial stages of attempts to use serological specificity for the fractionation of protein antigens have been described (Smith, Tozer, Gallop & Scanes, 1962). Individual antigen-rabbit antibody complexes were obtained from the interaction of mixtures ofantigens with mixtures of corresponding antibodies by using a modified gel-diffusion technique.…”