Large scale preparation of immunotoxins constructed with the Fab' fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain.
In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab' fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab' and dgA chain and the purification of the Fab'-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab'-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.
Ghetie
V
V
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
Ghetie
M A
MA
Uhr
J W
JW
Vitetta
E S
ES
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology*; D001709:Biotechnology / Q000379:methods; D015815:Cell Adhesion Molecules; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002845:Chromatography; D004731:Endotoxins / Q000032:analysis; D006023:Glycoproteins; D006863:Hydrogen-Ion Concentration; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D000922:Immunotoxins; D037102:Lectins; D010084:Oxidation-Reduction; D012276:Ricin; D051918:Sialic Acid Binding Ig-like Lectin 2
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D015815:Cell Adhesion Molecules; D004731:Endotoxins; D006023:Glycoproteins; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D000922:Immunotoxins; D037102:Lectins; D051918:Sialic Acid Binding Ig-like Lectin 2; D012276:Ricin