Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Antibodies, Monoclonal*
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Antigens, CD*
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Antigens, Differentiation, B-Lymphocyte / immunology*
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Biotechnology / methods
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Cell Adhesion Molecules*
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Chemical Phenomena
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Chemistry, Physical
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Chromatography
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Endotoxins / analysis
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Glycoproteins
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Hydrogen-Ion Concentration
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Immunoglobulin Fab Fragments
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Immunoglobulin G
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Immunotoxins*
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Lectins*
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Oxidation-Reduction
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Ricin*
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Sialic Acid Binding Ig-like Lectin 2
Substances
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Antibodies, Monoclonal
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Antigens, CD
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Antigens, Differentiation, B-Lymphocyte
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Cell Adhesion Molecules
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Endotoxins
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Glycoproteins
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Immunoglobulin Fab Fragments
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Immunoglobulin G
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Immunotoxins
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Lectins
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Sialic Acid Binding Ig-like Lectin 2
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Ricin