New article on optimizing in vitro protein expression purification

Cell-free protein expression strategies to optimize purification

By Douglas Hayworth

Our most recent product article describes strategies for optimizing expression and purification of functional proteins produced using the Thermo Scientificâ„¢ 1-Step High Yield IVT protein expression system.

The primary goal of this study was to learn if vectors with certain combinations of tags (and purification methods) would provide better overall results in terms of expression level and ease and quality of purification. Eric Hommema and collegues discovered that vectors containing an N- or C-terminal GST tag often provided greatly increased protein expression compared to vectors having only small antigen affinity tags. They also found that the multi-tag pT7CFE vectors containing both GST and His tags provided greater flexibility and success for purifications. The article includes complete protocols for both GST- and His-tag purification, as well as example data comparing results with different conditions.


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Choosing a vector and purification method for in vitro protein expression

Choosing a vector and purification method for in vitro protein expression