A novel "reverse screening" to identify refolding additives for activin-A.

Area of Science:

• Biochemistry
• Protein Chemistry
• Molecular Biology

Background:

• Refolding recombinant proteins from inclusion bodies (IBs) requires screening conditions to promote proper folding and minimize aggregation.
• Stabilizing native proteins and reducing aggregation are key strategies for successful refolding.

Purpose of the Study:

• To develop and evaluate a novel screening approach, termed reverse screening, for identifying effective protein refolding additives.
• To assess the efficacy of sodium taurodeoxycholate (TDCA) as a refolding additive for activin-A.

Main Methods:

• Partial unfolding of native activin-A in a buffer with urea and dithiothreitol.
• Monitoring protein recovery using reverse-phase high-performance chromatography.
• Testing various additives, including TDCA, for their ability to suppress aggregation and enhance recovery.

Main Results:

• Without additives, native activin-A experienced significant loss (approx. 60%) due to aggregation after 12-hour incubation.
• Sodium taurodeoxycholate (TDCA) substantially increased protein recovery and effectively suppressed aggregation.
• TDCA demonstrated high efficacy in refolding activin-A from inclusion bodies.

Conclusions:

• Reverse screening using native proteins is a viable approach for discovering effective refolding additives.
• TDCA is a highly effective additive for refolding proteins, including activin-A, from inclusion bodies.