NSF Award
9960806
This Small Business Innovation Research Phase I project will develop an enabling technology for computer-directed high throughput screening of proteins for improved properties. By combining a fast computer screen with experimental methods, the capabilities of random library screening can be greatly extended. This technology will be tested on Bacillus circulans xylanase with the goal of improving its activity at high temperature and pH. Starting from the enzyme crystal structure, Xencor's proprietary protein design automation (PDA) technology will be used to computationally prescreen the possible sequences resulting from varying selected residue positions, thereby eliminating unfavorable sequences and thus enormously reducing the number to be screened experimentally. The result of the computer screen is an amino acid probability<br/>distribution for the selected residue positions. A method will be developed to transform these probabilities into a DNA library that will be synthesized and experimentally screened for improved enzymatic activity. The computational prescreening will allow screening of ~10^80 sequences compared to 10^10 to 10^15 with conventional in vitro evolution techniques. The proposed research will result in a generally applicable<br/>procedure that allows optimization of far more residue positions and results in the more efficient development of improved proteins by coupling rational and random methods.
