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News & Blogs » Synthetic Biology News » Optimize Your Recombinant Protein Expression By Combinatorial Vector Design
An Introduction to Therapeutic Chimeric Antigen Receptor T Cells

High-Throughput DNA Library Assembly

Optimize Your Recombinant Protein Expression By Combinatorial Vector Design

Choosing and optimizing the best vector for protein or antibody expression could be complicated. There are many components that need to be taken into consideration while working on protein or antibody engineering and expression, including promoter and terminator choices, ribosomal binding site (RBS) sequences, signal peptide sequences, as well as different tag and linker options.

One traditional method used in protein expression is to optimize the expression system step by step, which takes several rounds of designing, cloning, and transfection. An alternative method would be to generate a panel of expression vectors with unique combinations of various components. Using this method, one can effectively synthesis different variants for each component and use advanced assembly technology to create a large number of constructs that covers all combinations, allowing the optimization of promoters, signal peptides, tags, linkers, as well as vector backbones in only one round. Therefore, the combinatorial method greatly minimizes the time and effort needed for the protein or antibody engineering and expression process.

Combinatorial modular assembly for recombinant protein and antibody expression.

Figure 1: Combinatorial modular assembly for recombinant protein and antibody expression.

By synthesizing each component variant and assemble desired combinations into one vector either randomly or individually, a large pool of protein expression modules can be constructed. Further screening and evaluation of all modules will help identify the best preforming expression vector for your protein or antibody.

Advantages of using the combinatorial method include:

  • Highly efficient than step-wise optimization;
  • High-throughput assembly for constructing every possible combination of your expression vector;
  • Ensures the identification of the most optimum construct for expressing your protein, where as step-wise optimization strategy can rarely identify the global optimum;
  • Recycle previous component variants for future projects without additional synthesis or cloning.

GenScript’s Combinatorial DNA Libraries

To facilitate the assembly of combinatorial libraries for protein and antibody expression, GenScript developed our own proprietary GenBuilder™ DNA assembly platform, which is able to assemble up to 12 parts in one round with insert length up to 15 Kb. Build upon this seamless assembly platform, GenScript now offers combinatorial DNA library services that can help speed up your recombinant protein and antibody engineering and expression process. Learn More »

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