Strategies for the depyrogenation of contaminated immunoglobulin G solutions by histidine-immobilized hollow fiber membrane.
The depyrogenation of different IgG solutions using the histidine-linked hollow fiber membrane developed in our laboratory is presented here. Three strategies for endotoxin (ET) removal were investigated according to the immobilized histidine's ability to bind different immunoglobulins: (1) ET removal from 1 mg/ml non histidine-binding mouse monoclonal IgG1 (MabCD4) solution was achieved in the presence of acetate buffer (pH 5.0) without any protein loss. (2) For contaminated human IgG, combined adsorption of ET and IgG in the presence of MOPS of Tris buffer was tested, followed by differential elution using increasing salt concentrations. This attempt was not successful since ET were quantitatively found in the IgG elution fraction. (3) Alternatively, it was proposed to adsorb selectively ET in the presence of acetate buffer (pH 5.0) under non binding conditions for human IgG. Human IgG could then be purified if necessary with the same membrane in the presence of MOPS buffer (pH 6.5). With a 1 m2 histidine-PEVA module under these operating conditions, it is estimated that the depyrogenation of 3 l of 1 mg/ml IgG (human or murine) solution containing 80 EU/ml of ET should be possible.
Legallais
C
C
Université de Technologie de Compiègne, Centre de recherches de Royallieu, Compiègne, France.
Anspach
F B
FB
Bueno
S M
SM
Haupt
K
K
Vijayalakshmi
M A
MA
D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods*; D004340:Drug Contamination; D004731:Endotoxins; D006639:Histidine; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008567:Membranes, Artificial; D051379:Mice; D011749:Pyrogens; D012996:Solutions
D000911:Antibodies, Monoclonal; D004731:Endotoxins; D007074:Immunoglobulin G; D008567:Membranes, Artificial; D011749:Pyrogens; D012996:Solutions; D006639:Histidine