Probing of some compounds as anti‐aggregatory additives in the protein refolding process from Escherichia coli inclusion bodies

Abstract

Five compounds of different chemical structure were tested for aggregation suppression during the refolding of porcine and mink growth hormones as model proteins from Escherichia coli inclusion bodies by the dilution method. Of all compounds tested in this work, 3-guanidinopropionic acid (GPA) containing a guanidinium group was the most effective additive for aggregation suppression. Anti-aggregatory properties of GPA were compared with the ones of l-arginine.

Bioscreening and expression of a camel anti-CTGF VHH nanobody and its renaturation by a novel dialysis–dilution method

“…During the dialysis method, the protein was retained in the dialysis bag and the proteins would start to refold while the denatured chemicals diffused outward. However, the dialysis was a complicated and time-consuming method requiring numerous repeats to optimize the refolding conditions and buffer composition, and generally taken multiple steps of dialysis (Jungbauer and Kaar 2007 ; Zilinskas and Sereikaite 2011 ; Berg et al 2012 ). As we known, during the protein refolding, the major obstruction came from the self-aggregation (Zilinskas and Sereikaite 2011 ; Gautam et al 2012 ).…”

“…However, the dialysis was a complicated and time-consuming method requiring numerous repeats to optimize the refolding conditions and buffer composition, and generally taken multiple steps of dialysis (Jungbauer and Kaar 2007 ; Zilinskas and Sereikaite 2011 ; Berg et al 2012 ). As we known, during the protein refolding, the major obstruction came from the self-aggregation (Zilinskas and Sereikaite 2011 ; Gautam et al 2012 ). Therefore, dilution was an effective method for in vitro protein refolding as it allowed the protein to refold at a very low concentration thus to reduce the interference from self-aggregation (Gautam et al 2012 ).…”

“…However, the dilution method was difficult to manipulate because the protein molecules needed to be separated into the solution instantly that required special equipment. Self-aggregation was also reported as the major obstacle for the dilution method in many cases (Zilinskas and Sereikaite 2011 ; Gautam et al 2012 ). To our surprising, the nanobody did not precipitate instantly after misfolded by normal dilution method but form small nanoparticles.…”

Growth hormone and growth hormone gene of the American mink (Neovison vison) – the current state of knowledge of one of the key hormones in one of the most intensively economically exploited species

“…It should be noted that a few studies on the mink somatotropin polypeptide are limited substantially to the development and improvement of methods for the obtaining the recombinant mGH in bacterial cells (Harada et al, 1994;Sereikaite et al, 2006;Sereikaite et al, 2007), as well as by optimizing the conditions of its storage and processing (Bajorunaite et al, 2007;Borromeo et al, 2008;Cirkovas, Sereikaite, 2010;Cirkovas, Sereikaite, 2011a,b;Zilinskas, Sereikaite 2011). This fact explains the scarcity of the available literature data on the specificity of the growth hormone molecule of N. vison (Sereikaite et al, 2006).…”