Three aspects of recombinant protein expression are key: quantity, purity and function. Many factors, such as poor sequence design and vector selection, contamination, suboptimal reagents and experimental conditions, will lead to protein insolubility, aggregation or misfolding, and ultimately affect the expression of recombinant proteins.
Strategy 1: Design expression vector
The strategy to avoid many of these problems is to design the best coding sequence and select the most appropriate expression vector for your specific downstream application.
Strategy 2: Increase protein solubility
Low protein solubility is the key factor to obtain low yield in recombinant expression. Before redesigning your sequence and expression vector, we suggest that you first operate some common expression condition parameters to improve protein solubility. Please click Download to view more details about condition parameters.
Strategy 3: Leverage Fusion Partners
Another strategy to optimize low yield protein expression is gene fusion chaperone technology. This strategy uses promoters, enhancers, tags and other fusion elements to increase the solubility of proteins.
Protein tags are short peptide sequences transplanted onto recombinant proteins, which are usually removed by enzymatic method or chemical reagents at the end of protein expression. Several common tags can be used in experiments to enhance protein expression. For more information about common tags, click Download.
Strategy 4: Improve purification
The last basic strategy is to improve your purity approach. Recombinant proteins can be purified manually or using a chromatographic system. Different purification methods can be planned according to the application and purity requirements you need. If the protein expression is stable, the one-step affinity purification method will produce enough protein and enough purity, which is suitable for various downstream applications. If further purification is required, size exclusion chromatography (SEC) can usually be performed after affinity purification. For further purification, ion exchange (IEX) chromatography can be used.