title: The GLP large scale preparation of immunotoxins containing deglycosylated ricin A chain and a hindered disulfide bond.
abstract: The large scale preparation of two second generation immunotoxins containing murine monoclonal antibodies and deglycosylated ricin A chain is described. The procedure for the preparation of immunotoxins consists of the derivatization of antibody with SMPT and reduction of dgA with DTT followed by their reaction to establish a hindered interchain disulfide bond. The purification of the immunotoxin includes affinity chromatography on Blue-Sepharose to remove the free antibody and gel filtration on Sephacryl S-200HR to remove any high molecular weight material and free dgA. The two immunotoxins were prepared by GLP procedures and tested for yield, composition, purity, sterility and biological activity.
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
authors.forename: V
authors.initials: V
authors.lastname: Ghetie
authors.forename: P
authors.initials: P
authors.lastname: Thorpe
authors.forename: M A
authors.initials: MA
authors.lastname: Ghetie
authors.forename: P
authors.initials: P
authors.lastname: Knowles
authors.forename: J W
authors.initials: JW
authors.lastname: Uhr
authors.forename: E S
authors.initials: ES
authors.lastname: Vitetta
mesh_terms: D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000276:immunology; D018941:Antigens, CD19; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology; D015815:Cell Adhesion Molecules; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D003545:Cysteine; D004591:Electrophoresis, Polyacrylamide Gel; D000922:Immunotoxins; D037102:Lectins; D012276:Ricin; D051918:Sialic Acid Binding Ig-like Lectin 2; D013388:Succinimides
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018941:Antigens, CD19; D000944:Antigens, Differentiation, B-Lymphocyte; D015815:Cell Adhesion Molecules; D003432:Cross-Linking Reagents; D000922:Immunotoxins; D037102:Lectins; D051918:Sialic Acid Binding Ig-like Lectin 2; D013388:Succinimides; C058151:4-succinimidyloxycarbonyl-alpha-methyl-alpha(2-pyridyldithio)toluene; D012276:Ricin; D003545:Cysteine