Proprietary library-based screening technology to identify solubly expressing constructs of problematic proteins
Expression of soluble protein by random incremental truncation
- The ESPRIT construct screening platform has been developed at EMBL to identify soluble constructs of "difficult-to-express" protein targets that resist the classical approach of bioinformatics and PCR cloning.
It employs the principles of directed evolution whereby a diverse random library of DNA constructs is generated and screened to identify rare clones of interest (soluble expressers).
All unidirectional truncations of the target gene, both 5’ and 3’, are synthesised by exonuclease degradation to generate potential expression constructs.
Additionally, we have developed a "scanning" version to identify internal domains. In each experiment, 30,000 individual clones are assayed in parallel for yield and solubility using a highly automated colony array format.