Purification and properties of immunotoxins containing one vs. two deglycosylated ricin A chains.
A method for the preparation of immunotoxins (ITs) containing one and two molecules of deglycosylated ricin A chain (dgA) bound to each molecule of anti-human CD22 mouse IgG1 monoclonal antibody is described. The cross-linking of antibody and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-alpha-methyl-(2-pyridyldithio) toluene. The two ITs were purified by successive chromatographies on Blue-Sepharose, Sephacryl S-200, Blue-Sepharose with NaCl gradient and Sephacryl S-300. The IT with two dgAs was seven-fold more cytotoxic to CD22+ Daudi cells than that containing one molecule of dgA.
Ghetie
V
V
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235-9048.
Swindell
E
E
Uhr
J W
JW
Vitetta
E S
ES
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000633:toxicity; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D015815:Cell Adhesion Molecules; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D000922:Immunotoxins / Q000302:isolation & purification* / Q000633:toxicity; D037102:Lectins; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D012276:Ricin / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D051918:Sialic Acid Binding Ig-like Lectin 2
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C497867:CD22 protein, human; C497868:Cd22 protein, mouse; D015815:Cell Adhesion Molecules; D007074:Immunoglobulin G; D000922:Immunotoxins; D037102:Lectins; D051918:Sialic Acid Binding Ig-like Lectin 2; D012276:Ricin