Elastin protein purification

Wu WY, Fong BA, Gilles AG, Wood DW (2009) Recombinant protein purification by selfcleaving elastin-like polypeptide fusion tag. Curr Protoc Prot Sci 26.4 21-18... 

Fig. 11.3 Purification ofSOl-lOOxELP-proteins from transgenic tobacco plants by inverse transition cycling and analysis by SDS-PAGE. 1 15 pg of total soluble leaf protein extracted in raw extract buffer 2 cleared supernatant of original 15 pg total soluble leaf protein after heat treatment (60 min, 95 °C) 3 cleared supernatant of original 300 pg leaf protein after heat treatment 4 cleared supernatant of original 300 pg leaf protein after heat treatment (60 min, 60 °C) with 2 M NaCI 5 redissolved spider silk-elastin protein pellet from original 300 pg of total soluble leaf protein after heat treatment (60 min, 60 °C) with 2 M NaCI.

Meyer DE, Trabbic-Carlson K, Chilkoti A (2001) Protein purification by fusion with an environmentally responsive elastin-fike polypeptide effect of polypeptide length on the... 

In addition to the commonly used synthetic polymers such as pNIPAM and poly(N-vinylcaprolactam) (PVCL), elastin-like polymers were introduced as thermoresponsive polymers for affinity precipitation (Shimazu et al., 2003). An elastin-like polymer consists of the repeating penta-peptide, Val-Pro-Gly-Xaa-Gly (Xaa being any amino acid except proline). It undergoes reversible phase transition under temperature or salt concentration changes similar to pNIPAM (Shimazu et a/., 2003). An elastin-like polymer was used as a terminal tag for the facilitation of recombinant protein purification as attaching it to the hydrolase enzyme improves its purification. Purification, in excess of 1300-fold was achieved after two cycles of reversible changes of elastin-like polypeptide conformation precipitation induced by environmental changes (Shimazu et al., 2003). 

Elastin-like polypeptides (ELPs) are thermoresponsive biopolymers with a broad range of biotechnological applications including tissue engineering, drug delivery, biosensing, and protein purification [118, 119]. like PNIPAAm, ELPs also... 

Purification of Spider Silk-Elastin Fusion Proteins by Heat Treatment and Inverse Transition Cycling... 

Figure 3 Biosynthesis and purification of 90-kD elastin analogue analyzed by denaturing polyacrylamide gel electrophoresis (10-15% gradient, visualized by silver staining). Lanes 1-7 time course of target protein expression at 0, 30, 60, 90, 120, 150, and 180 minutes after induction. Lane 9 soluble lysate of induced E. coli expression strain BLR(DE3)pRAMl. Lanes 10-13 protein fractions obtained from immobilized metal affinity chromatography of the lysate on nickel-NTA agarose (imidazole gradient elution). Lanes 8,14 protein molecular weight standards of 50, 75, 100, and 150 kD.

Sandberg, L. B., Weissman, N., and Smith, D. W. The purification and partial characterization of a soluble elastin-like protein from copper-deficient porcine aorta. Biochemistry 8, 2940-2945 (1969). 

Conley AJ, Joensuu JJ, Jevnikar AM, Menassa R, Biandle JE (2009) Optimization of elastin-like polypeptide fusions for expression and purification of recombinant proteins in plants. Biotechnol Bioeng 103 562-573... 

Lim DW, Trabbic-Carlson K, Mackay JA, Chilkoti A (2007) Improved non-chromatographic purification of a recombinant protein by cationic elastin-fike polypeptides. Biomacromolecules 8 1417-1424... 

Other proteins such as elastin, silk fibroin, keratin or albumin have been considered for materials applications very recently. Due to the lack of availability or other purification and processing capacities, their cost remains relatively high. Consequently, up to now, their main material applications concern the high added value materials, mainly biomedical. 

Tlie elastin-like polypeptides have been also tested as carriers for metalbinding ligands (Stiborova e/ai,2003).The synthesised elastin-like polymer was chemically modified by introducing imidazole functionality. Purification of His-tag proteins, ]8-D-galactose and chloramphenicol acetyl transferase was performed in a manner similar to pNIPAM-vinyl imidazole systems. 

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