The major antigenic beer macromolecule, Antigen 1, has been isolated from commercial lager beer by succesive use of ultrafiltration, alcohol precipitation, anion exchange, cation exchange and gel filtration chromatography. Amino acid analyses showed a composition identical with that of a major barley albumin, Protein Z, except for a 16% lower content of lysine. Like Protein Z, Antigen 1 showed a molecular weight (MW) near 40,000 in gel filtration and SDS‐gel electrophoresis. Unlike Protein Z, Antigen 1 contained 2·5% carbohydrate and was more acidic. When properly standardised, the amount of Antigen 1 in beer could be determined immunoelectrophoretically. Contents ranged from 22 mg/litre in a pale ale (7·7°P; 36% brewers adjunct) to 170mg/litre in a stout (18·8°P). The results suggest that Antigen 1 may account for more than 10% of the total non‐dialysable proteinaceous material in beer, and that about 25% of the Protein Z present in the brewing materials may be recovered with an almost unmodified protein structure as Antigen 1 of beer. Apparently, Antigen 1 was not affected by stabilisation of the beer with insoluble PVP or with papaya proteinases.

The macromolecules of an antigen-rich beer fraction X were characterized by column chromatography. Elution profiles of protein, carbohydrate and immunochemicaliy distinct antigens were recorded after separation on gels of Sephacryl, Sephadex, Sepharose, DEAE-Sephadex, SPSephadex, Bio-gel HT hydroxylapatite. Con A-Sepharose, and two types of antibody-Sepharose. The elution patterns demonstrate that beer antigens, originating from barley or yeast macromolecules, can be separated from carbohydrates, from 'proteose-like' material and from each other in a few preparative chromatographic steps. Affinity chromatography on a column of im mobilized antibodies towards fraction X indicated that the antigenic material constitutes about 25% of the fraction. More than 20% of the protein in fraction X seems to originate from a distinct barley protein Z.

Very-high-purity, double virus-inactivated vWf preparations containing high molecular weight multimers were obtained on a production scale.

No abstract

Background and Objectives: We investigated large–scale production of very–high–purity von Willebrand factor (vWf) containing high–molecular–weight vWf multimers, using chromatography alone. Materials and Methods: Factor VIII (FVIII)–containing vWf was produced by initial separation of vWf from plasma by gel filtration followed by two ion exchange steps. Two virus inactivation steps were incorporated in the process. Results: The process resulted in FVIII–containing vWf preparations with a mean specific activity of 82 U vWf: collagen–binding activity per milligram protein (excluding added albumin) and with almost intact vWf multimer distributions. Conclusions: Very–high–purity, double virus–inactivated vWf preparations containing high molecular weight multimers were obtained on a production scale.