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With breakthroughs in DNA synthesis, high-throughput scanning, and protein engineering, directed evolution (DE) can replace the process of long-term natural selection by precisely engineering proteins and/or nucleic acids toward a user-defined feature. Directed evolution has been widely used in the optimization and analysis of DNA sequences, gene function, and protein structure. The process of directed evolution includes generation of random gene libraries, gene expression in suitable hosts, and screening of mutant libraries. The key to construct a gene library is its capacity and mutation diversity, while the key to screening variants is high sensitivity and high-throughput. Synbio Technologies commits to DNA solutions to DE by building both sensitive screening and cost-effective DNA library variants ranging from alanine scan libraries, site saturation libraries, modular substitution libraries, trimer libraries, and antibody libraries (Tab.1).
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| Library Types
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Description |
Schema
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| Alanine Scan Library
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Research on amino acid residues critical to protein function, interaction, and shape by replacing an individual amino acid with alanine at every position.
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| Site Saturation Library |
Substitute all 20 possible amino acids at any site to evaluate structure-function relationships and generate improved protein variants.
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| Antibody Library |
Create a high-diversity synthetic antibody library or improve existing antibody functionality such as specificity, immunogenicity, affinity, expression, and/or aggregation.
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| Modular Substitution Library |
Build new genetic modules, regions, or pathways from DNA parts including regulatory elements and/or coding sequences.
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| Trimer Library |
Introduce 20 amino acids into the specific sites of proteins by accurately control the type and proportion of amino acids through chemically synthesized trimer oligos.
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| | Tab.1 The type of synthetic DNA libraries |
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Specific protein-protein interaction (PPI) is the core function of protein, and improving PPI and even obtaining new PPI through directed evolution is an important goal of protein engineering. Our ntibody design and production platform can mimic the natural evolution of protein-protein interaction that significantly powers the discovery of ideal protein such as an antibody. Directed evolution of proteins has been successfully applied in the design of key enzymes in metabolic pathways, the creation of novel functional proteins, and the screening and identification of expected functional proteins. It has played an important role in the fields of metabolic engineering and synthetic biology.
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Case Study
Fast Discovery of High Affinity PDL1 Antibody Through DE
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De novo antibody design and affinity evolution
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High binding affinity was obtained through DE approach
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| Loading Sample ID |
kon(1/Ms) |
kdis(1/s) |
KD (M) |
Full R^2 |
| 55_56 |
9.28E+03 |
2.70E-07 |
2.91E-11 |
0.992429 |
| 57_58 |
3.76E+03 |
5.60E-07 |
1.49E-10 |
0.995068 |
| Positive control |
7.29E+04 |
8.90E-07 |
1.22E-11 |
0.996965 |
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