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.
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