Evaluation of IDA-PEVA hollow fiber membrane metal ion affinity chromatography for purification of a histidine-tagged human proinsulin.
Inabilities to process particulate material and to allow the use of high flow rates are limitations of conventional chromatography. Membranes have been suggested as matrix for affinity separation due to advantages such as allowing high flow rates and low-pressure drops. This work evaluated the feasibility of using an iminodiacetic acid linked poly(ethylenevinyl alcohol) membrane in the immobilized metal ion affinity chromatography (IMAC) purification of a human proinsulin(His)(6) of an industrial insulin production process. The screening of metal ions showed Ni(2+) as metal with higher selectivity and capacity among the Cu(2+), Ni(2+), Zn(2+) and Co(2+). The membrane showed to be equivalent to conventional chelating beads in terms of selectivity and had a lower capacity (3.68 mg/g versus 12.26 mg/g). The dynamic adsorption capacity for human proinsulin(His)(6) was unaffected by the mode of operation (dead-end and cross-flow filtration).
Department of Bioprocess Engineering, School of Chemical Engineering, State University of Campinas, UNICAMP, CP 6066, CEP 13083-970, Campinas, SP, Brazil.
Luciana Cristina Lins
LC
de Aquino
Heloisa Ribeiro Tunes
HR
de Sousa
Everson Alves
EA
Miranda
Luciano
L
Vilela
Sônia Maria Alves
SM
Bueno
D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D008567:Membranes, Artificial; D008670:Metals / Q000737:chemistry*; D011384:Proinsulin / Q000737:chemistry / Q000302:isolation & purification*; D013816:Thermodynamics
D008567:Membranes, Artificial; D008670:Metals; D006639:Histidine; D011384:Proinsulin