One-step purification of histidine-tagged profilin with high purity and yield by using metal precipitation.
A simple one-step method for the purification of recombinant His-tagged profilin from the bacterial cell lysate is reported. Noting the greater ease with which hexahistidine-tagged proteins can be metalprecipitated compared with unwanted protein impurities, we investigated the effect of lysis-buffer additives and optimization of other conditions to recover selectively desired proteins in a one-step metal precipitation without using biopolymers. Purification of the His-tagged melon (Cucumis melo) profilin was used to demonstrate the utility of this method and up to 80% recovery with a purity of 98% was achieved. This method obtained a yield of the protein nearly comparable with that obtained using metal-affinity chromatography. This purification procedure can reduce the time and cost of the purification process, especially on a large scale.
Sankian
Mojtaba
M
Immunobiochemistry Laboratory, Immunology Research Center, Bu-Ali Research Institute, Ferdossi Square, Bu-Ali Square, Mashhad, Mashhad, Iran.
Yousefi
Mitra
M
Pazouki
Nazanin
N
Varasteh
Abdolreza
A
D000345:Affinity Labels / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008670:Metals / Q000737:chemistry; D051304:Profilins / Q000737:chemistry* / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
D000345:Affinity Labels; D008670:Metals; D051304:Profilins; D011994:Recombinant Proteins