Batch affinity adsorption of His-tagged proteins with EDTA-based chitosan

Abstract

Affinity adsorption purification of hexahistidine-tagged (His-tagged) proteins using EDTA-chitosan-based adsorption was designed and carried out. Chitosan was elaborated with ethylenediaminetetraacetic acid (EDTA), and the resulting polymer was characterized by FTIR, TGA, and TEM. Different metals including Ni(2+), Cu(2+), and Zn(2+) were immobilized with EDTA-chitosan, and their capability to the specific adsorption of His-tagged proteins were then investigated. The results showed that Ni(2+)-EDTA-chitosan and Zn(2+)-EDTA-chitosan had high affinity toward the His-tagged proteins, thus isolating them from protein mixture. The target fluorescent-labeled hexahistidine protein remained its fluorescent characteristic throughout the purification procedure when Zn(2+)-EDTA-chitosan was used as a sorbent, wherein the real-time monitor was performed to examine the immigration of fluorescent-labeled His-tagged protein. Comparatively, Zn(2+)-EDTA-chitosan showed more specific binding ability for the target protein, but with less binding capacity. It was further proved that this purification system could be recovered and reused at least for 5 times and could run on large scales. The presented M(2+)-EDTA-chitosan system, with the capability to specifically bind His-tagged proteins, make the purification of His-tagged proteins easy to handle, leaving out fussy preliminary treatment, and with the possibility of continuous processing and a reduction in operational cost in relation to the costs of conventional processes.

Synthesis of a bifunctional EDTA–carboxymethyl chitosan derivative and its potential as an adsorbent for the removal of Cu2+ ions from aqueous solutions

“…In addition, the broadband around 3367 cm À1 attributes to the O-H stretching vibrations along with the overlapped N-H stretching vibrations. 27 The characteristic C-O and C-O-C stretching vibrations of the polysaccharide chain of CMC are detected in the range of 1150-897 cm À1 (Figure 2(E)) and those are similar to the vibrations observed for chitosan. The characteristic vibrations observed at 1730 cm À1 and 1249 cm À1 are assigned to the symmetric stretching vibration of C=O in COOH group and stretching vibration of C-O in CH 2 COOH, respectively, that cannot be found in chitosan.…”

“…The second stage of weight loss for chitosan occur around 300°C due to scission of ether linkages in chitosan backbone. 3,13,27,29 Similar to above, degradation of polymer causes a weight loss around 210°C for CMC. In EDTA-CMC, degradation of polymer along with different functional groups corresponds to a weight loss around 225°C.…”

“…In addition, the broadband around 3367 cm −1 attributes to the O-H stretching vibrations along with the overlapped N-H stretching vibrations. 27

Figure 2.FT-IR spectra of chitosan (C) and CMC (E).

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Synergistic effects of EDTA and lysozyme on the properties of hydroxypropyl starch nano antibacterial films

“…was more sensitive to the synergistic inhibitory effect of EDTA and LY, while S. aureus was less susceptible to the synergistic inhibition. According to literature, EDTA has a strong antibacterial activity on GNB and a weaker antibacterial activity on GPB( Hua et al, 2016 ). Simultaneously, as an attractive antibacterial agent in the food and medical care industry, the antibacterial activity of LY was related to its muramidase activity, cationic and hydrophobicity ( Masschalck and Michiels 2003 ).…”

Synthesis of human hair fiber-impregnated chitosan beads functionalized with citric acid for the adsorption of lysozyme

“…29 Hua et al modied chitosan with ethylenediaminetetraacetic acid and the obtained materials could specically bind his-tagged proteins. 30 Sun et al synthesized carboxymethyl chitosan magnetic nanoparticles by a chemical coprecipitation method and then treated these with various ion solutions for lysozyme separation and purication. 31 Recently, Ding et al have demonstrated another fascinating protein adsorbent by in situ functionalizing electrospun ethylene-vinyl alcohol nanobrous membranes with critic acid (CA).…”