Evaluation of immobilized metal membrane affinity chromatography for purification of an immunoglobulin G1 monoclonal antibody.
The large scale production of monoclonal antibodies (McAbs) has gaining increased relevance with the development of the hybridoma cell culture in bioreactors creating a need for specific efficient bioseparation techniques. Conventional fixed bead affinity adsorption commonly applied for McAbs purification has the drawback of low flow rates and colmatage. We developed and evaluated a immobilized metal affinity chromatographies (IMAC) affinity membrane for the purification of anti-TNP IgG(1) mouse McAbs. We immobilized metal ions on a poly(ethylene vinyl alcohol) hollow fiber membrane (Me(2+)-IDA-PEVA) and applied it for the purification of this McAbs from cell culture supernatant after precipitation with 50% saturation of ammonium sulphate. The purity of IgG(1) in the eluate fractions was high when eluted from Zn(2+) complex. The anti-TNP antibody could be eluted under conditions causing no loss of antigen binding capacity. The purification procedure can be considered as an alternative to the biospecific adsorbent commonly applied for mouse IgG(1) purification, the protein G-Sepharose.
Serpa
Gisele
G
Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química, UNICAMP, CP 6606, CEP 13083-970, Campinas, SP, Brazil.
Augusto
Elisabeth Fátima Pires
EF
Tamashiro
Wirla Maria Silva Cunha
WM
Ribeiro
Mariana Borçoe
MB
Miranda
Everson Alves
EA
Bueno
Sônia Maria Alves
SM
D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008024:Ligands; D008567:Membranes, Artificial; D008670:Metals; D051379:Mice; D013816:Thermodynamics; D015032:Zinc
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008024:Ligands; D008567:Membranes, Artificial; D008670:Metals; D015032:Zinc