Microwave assisted solubilization of inclusion bodies

Sustainable Chemical Processes

Table 2 Effect of microwave treatment on the average sizes of different inclusion bodies dissolved in different denaturants

Inclusion Bodies	Average diameter (nm) of inclusion bodies in buffer before microwave treatment	Average diameter (nm) after microwave treatment.
		8 M urea	6 M GuHCl	20 mM SDS
MBP224D	1493	390	451	130
MBP264D	1504	440	326	130
CcdB-M97K	751	93	438	62
CD4D12	1040	284	338	295
MalETrx	731	120	410	195
ScFv	1444	615	587	560
ScFab	1400	884	811	547

Inclusion bodies of MBP224D, MBP264D, CcdB-M97K, CD4D12, malETrx, ScFv and ScFab, were dissolved in 8 M urea, 6 M GuHCl and 20 mM SDS and subjected to microwave treatment for 2 minutes and the average diameter was recorded. The average diameters of inclusion bodies suspended in 50 mM Tris–HCl, pH 7.0 (without denaturant) and prior to microwave treatment were also recorded for comparison. The average sizes of the dissolved inclusion bodies were measured by dynamic light scattering. In all the measurements the buffer used was 50 mM Tris–HCl, pH 7.0. The temperature during microwave solubilization was maintained at 26 ± 1°C. Protein concentration in all the cases was 2.0 mg.mL^-1. The readings were taken in triplicates and the difference among the individual readings was less than 5%.