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.
- Fab′ fragment
- Large scale preparation of proteins
- Monoclonal antibody
- Ricin toxin deglycosylated A chain
ASJC Scopus subject areas
- Immunology and Allergy