Negative chromatography on agarose-TREN as a technique for purification of protein spiked in soybean seeds extract

作者：Antunes Pereira Bresolin Iara Rocha; Lazzarotto Bresolin Igor Tadeu; Miranda Everson Alves; Alves Bueno Sonia Maria

来源：Process Biochemistry, 2012, 47(12): 2255-2261.

DOI：10.1016/j.procbio.2012.08.023

• Tris(2-aminoethyl)amine (TREN)
• Mixed-mode chromatography
• Negative chromatography
• Soybean extract
• Downstream processing

Alkyl amines and polyamines have been used as ligands for protein purification by mixed-mode chromatography. The adsorption of proteins onto these ligands seems to be governed by multiple effects such as electrostatic, hydrophobic, and affinity interactions. In this work we investigated the adsorption of proteins extracted from soybean onto the adsorbent agarose-Tris(2-aminoethyl)amine (TREN). The effects of flow rate, buffer system, and extract concentration on the capture of proteins extracted from soybean were evaluated. Experiments using Mes at pH 6.5 as adsorption buffer allowed the adsorption of almost the totality of native soybean protein with a dynamic adsorption capacity of 13.50 mg mL(-1) adsorbent. Experiments with human IgG (pl in the range of 5.8-9.0) and human serum albumin (HSA, pl of 4.9) spiked into these extracts lead to the conclusion that electrostatic forces play a major role in the interaction between protein and agarose-TREN. Based on this work, negative chromatography with agarose-TREN should be considered as a method for purification of basic recombinant protein produced in transgenic soybean seeds.

• 出版日期2012-12