Isolation of Antibodies to Gliadin and Ragweed Antigen E with the Use of Immunosorbents

Adsorption of Human Reagins to Ethylchloroformate Polymerized Antigen E and Desorption with Thiocyanate Ions

“…The present study has confirmed the finding of G oldstein et al [5] that ethyl chloroformate-polymerized protein allergens can function as specific immunoadsorbents for the separation of allergen-binding anti bodies, including reagins, from human atopic serum. In this investiga tion, attention was specifically directed to the separation of reaginic anti bodies in ragweed atopic serum by immunoadsorption with polymerized antigen E. The P-K test was adopted as the method of assay to deter mine the extent of adsorption and desorption of these antibodies, under the assumption that the P-K titers obtained were directly proportional to reaginic antibody content.…”

“…According to the data of G oldstein et al [5], some 20-40°/o of the reaginic antibodies in ragweed atopic serum were adsorbed to polymer ized antigen E under the specific conditions of incubation employed in their study (30-min incubation with a 40 mg polymer). Similar levels of adsorption (20°/o) were observed in the present study under the same conditions ( fig.…”

“…Reagins (as well as hemagglutinins) to ragweed or gliadin were desorbed from the insoluble allergen preparations by in cubation with buffered salt solutions of high ionic strength, viz., 2 m NaCl or 2 m KI. However, the yields obtained for both types of reaginic antibodies were very low [5]. In the present study, an attempt was made to improve upon the yields of ragweed-specific reagins by the develop-ment of more optimal conditions for adsorption to and elution from po lymerized antigen E. It was of particular interest to determine if thio cyanate would prove superior to chloride or iodide for elution of the ad sorbed reagins in view of the demonstration by D andliker et al [2] that, compared to other chaotropic ions, thiocyanate was most effica cious for dissociation of rabbit antibodies from specific immunoadsorbents.…”

“…Insoluble antigen E immunoadsorbent was prepared by polymerizing antigen E according to the method previously described [5] with minor modifications. 40 mg of antigen E were dissolved in 5 ml of 0.005 m NaHCOs and the solution brought to pH 5.0 with the addition of 2 ml of 0.02 m sodium acetate buffer, pH 5.0.…”

“…In a previous study in this laboratory, Goldstein et al [5] applied the immunoadsorption method of A vrameas and T ernynk [1] to sepa rate reaginic antibodies from wheat and ragweed sensitive human sera. Gliadin and ragweed antigen E were individually copolymerized with ethyl chloroformate and the insoluble polymers incubated with the cor responding allergic sera.…”