The GLP large scale preparation of immunotoxins containing deglycosylated ricin A chain and a hindered disulfide bond.
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.
Ghetie
V
V
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
Thorpe
P
P
Ghetie
M A
MA
Knowles
P
P
Uhr
J W
JW
Vitetta
E S
ES
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
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