Table 1 Solubilization and refolding of proteins of interest (POIs).

Protein	Stabilization buffer	Refolding buffer		Calculated pI	tES charge	Diameter of tES encapsulated protein (nm)
Protein	Stabilization buffer	Buffer	Additive	Calculated pI	tES charge	TEM	DLS
α conotoxin	S + 2 mM EDTA*	R₂	NA	7.8	Negative	13.14 ± 1.01	11.78 ± 1.19
λ conotoxin	S + 2 mM EDTA*	R₂	NA	8	Neutral	11.41 ± 1.53	11.55 ± 0.76
rFasxiator	S	R₁	+ 1 mM GSSG/ 0.8 mM GSH/5 mM CaCl₂	8.3	Negative	13.13 ± 1.23	11.73 ± 0.73
PLA₂	S*	R₁	+ 4 mM GSSG/2 mM cysteine/10 mM CaCl₂	4.8	Positive	12.60 ± 1.39	12.27 ± 0.68
GFPuv	S*	R₁	NA	5.8	Positive	13.10 ± 1.12	12.23 ± 0.28
HRPc	S*	R₁	+ 0.35 mM GSSG/5 mM CaCl₂/20 uM heme	6.3	Positive	13.16 ± 1,06	11.71 ± 0.86
rLuc	S*	R₁	NA	5.86	Positive	12.87 ± 1.33	11.17 ± 0.79
FFL	S	R₁	+ 50 mM KCl/ 3 mM MgCl₂/1 mM DTT	6.69	Neutral	13.05 ± 1.23	11.53 ± 1.17
HSA	S	R₁	+ 2 mM GSSG/1 mM GSH/1 mM EDTA	5.67	Positive	11. 99 ± 1.42	12.08 ± 1.24
sAP	S	R₁	+ 0.8 mM GSSG/ 1 mM GSH/2 mM + MgCl₂/4 mM ZnCl₂	4.63	Positive	13.23 ± 1.15	NA
Omp2a	S	R₁	+ 0.005% Triton-X	4.3	Positive	12.27 ± 2.06	NA
p53	S	R1	(a)2 mM DTT/ 1 mM ZnCl₂/(b) 1 mMZncl2	6.33	Neutral	12.45 ± 1.46	NA

S: 50 mM Tris-Hcl - pH 8.0, 8 M Urea (or 6 M GuHcl), 0.1-0.2 M NaCl, 5 mM DTT (or 10 mM BME), *Heat at 60 °C for 30 min, R1: 50–100 mM Tris-Hcl - pH 8.0, 50-400 mM NaCl, 1–7% Glycerol, R2: 100 mM Tris-Hcl - pH 8.0, 2 mM EDTA, 1 mM GSSG,0.8 mM GSH, PLA₂: Phospholipase A₂, GFPuv: Green Fluorescent Protein, HRPc: Horseradish Peroxidase, rLuc: Renilla Luciferase, FFL: Firefly Luciferase, HSA: Human Serum Albumin, sAP: Shrimp Alkaline Phosphotase. Details regarding solubilization buffer, refolding buffer, pI of POI, tES charge, diameter of tES:POI assembly analysed by DLS and TEM.

Quick links

Search