caa a22 4500 465734316 CHVBK 20180323111750.0 cr unu---uuuuu 170327e19901101xx s 000 0 eng 10.1007/BF01189079 doi (NATIONALLICENCE)springer-10.1007/BF01189079 Immobilization of galactosyltransferase and continuous galactosylation of glycoproteins in a reactor [Elektronische Daten] [Robert Schneider, Mario Hammel, Eric Berger, Oreste Ghisalba, Jakob Nueesch, Daniel Gygax] We immobilized human milk galactosyltransferase covalently to CNBr- and tresylchloride-activated Sepharose. The enzyme was also immobilized non-covalently to Concanavalin A-Sepharose and to monoclonal anti-galactosyltransferase antibodies which were boundvia their Fc-fragment to Protein G-Sepharose. With the covalent methods, up to 72% of the enzyme could be bound to the carrier, but more than 90% of the specific activity was lost. In contrast, non-covalent immobilization yielded only about 50% immobilization efficiency, but 21% and 25% of specific activity, respectively, could be recovered. The stability of immobilized galactosyltransferase as compared to native enzyme was considerably increased: at room temperature, 55% of initial immobilized activity was lost after 65 hours compared to 95% of loss of soluble enzyme activity. Immobilized galactosyltransferase was then used for continuous galactosylation of the glycoproteins ovalbumin, endo H-treated yeast invertase and bovine serum albumin-N-acetylglucosamine in a "slurry” reactor. 55%, 35% and 25%, respectively, of all acceptor sites on these glycoproteins could be galactosylated by this method. Glycoconjugate Journal, 1990 glycosylation nationallicence glycosyltransferases nationallicence galactosyltransferase nationallicence recombinant glycoproteins nationallicence enzyme immobilization nationallicence enzyme reactor nationallicence Schneider Robert Central Research Laboratories, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut Hammel Mario Central Research Laboratories, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut Berger Eric Institute of Physiology, University of Zurich, Winterthurerstr. 190, 8057, Zurich, Switzerland aut Ghisalba Oreste Central Research Laboratories, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut Nueesch Jakob Pharma Division, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut Gygax Daniel Pharma Division, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut Glycoconjugate Journal Kluwer Academic Publishers 7/6(1990-11-01), 589-600 0282-0080 7:6<589 1990 7 10719 https://doi.org/10.1007/BF01189079 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01189079 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Schneider Robert Central Research Laboratories, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut NATIONALLICENCE 700 1- Hammel Mario Central Research Laboratories, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut NATIONALLICENCE 700 1- Berger Eric Institute of Physiology, University of Zurich, Winterthurerstr. 190, 8057, Zurich, Switzerland aut NATIONALLICENCE 700 1- Ghisalba Oreste Central Research Laboratories, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut NATIONALLICENCE 700 1- Nueesch Jakob Pharma Division, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut NATIONALLICENCE 700 1- Gygax Daniel Pharma Division, Ciba-Geigy AG, Klybeckstr. 141, 4057, Basel, Switzerland aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 7/6(1990-11-01), 589-600 0282-0080 7:6<589 1990 7 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer