Protein purification with polymeric affinity membranes containing functionalized poly(acid) brushes.
Porous nylon membranes modified with poly(acid) brushes and their derivatives can rapidly purify proteins via ion-exchange and metal-ion affinity interactions. Membranes containing poly(2-(methacryloyloxy)ethyl succinate) (poly(MES)) brushes bind 118 +/- 8 mg of lysozyme per cm(3) of membrane and facilitate purification of lysozyme from chicken egg white. Moreover, functionalization of the poly(MES) brushes with nitrilotriacetate (NTA)-Ni(2+) complexes yields membranes that bind poly(histidine)-tagged (His-tagged) ubiquitin with a capacity of 85 +/- 2 mg of protein per cm(3) of membrane. Most importantly, the membranes modified with poly(MES)-NTA-Ni(2+) allow isolation of His-tagged cellular retinaldehyde-binding protein directly from a cell extract in <10 min, and the protein purity is comparable to that achieved with commercial affinity columns. Therefore, porous nylon membranes containing functionalized poly(MES) brushes are attractive candidates for rapid, high-capacity purification of His-tagged proteins from cell extracts.
Jain
Parul
P
Department of Chemistry Michigan State University East Lansing, Michigan 48824, USA.
Vyas
Mukesh Kumar
MK
Geiger
James H
JH
Baker
Gregory L
GL
Bruening
Merlin L
ML
D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D004529:Egg White / Q000737:chemistry; D006639:Histidine / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008567:Membranes, Artificial; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009532:Nickel / Q000737:chemistry / Q000378:metabolism; D009571:Nitrilotriacetic Acid / Q000737:chemistry* / Q000378:metabolism; D009757:Nylons / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017550:Spectroscopy, Fourier Transform Infrared; D025801:Ubiquitin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
C020932:11-cis-retinal-binding protein; D002352:Carrier Proteins; D008567:Membranes, Artificial; D009757:Nylons; D011108:Polymers; D025801:Ubiquitin; D006639:Histidine; D009532:Nickel; D009113:Muramidase; D009571:Nitrilotriacetic Acid