Scanning Point Mutation Library Construction

Scanning Point Mutations improves the standard alanine/cysteine scanning by substituting each amino acid with all 20 amino acids simultaneously. The technique is a means of improving the properties of proteins and can provide details of each amino acid at that position. For each desired codon, a small site-saturation library can be constructed.

Alanine (Ala) scanning is a widely used mutagenesis method in which amino acid residues in a target protein are systematically substituted with alanine at selected positions. Studies have shown that substitution with alanine can eliminate side-chain interactions without changing the main-chain conformation or introducing steric and electrostatic effects, so it is often used to test specific side-chain contributions while preserving the native protein structure.

Alanine scanning libraries can be used to identify specific amino acid residues responsible for peptide stability, function, and conformation. During the experiment, alanine was sequentially substituted for each non-alanine residue one at a time, and the corresponding change in epitope activity was measured. This library allows rapid determination of the contribution of each amino acid's contribution to the peptide's functionality.

Figure 1. Shotgun scanning (Morrison, K.L.; Weiss, G.A. 2001)

Services

As an expert in protein research, Profacgen provides comprehensive alanine scanning mutagenesis and high-throughput analysis to help customers with epitope mapping and protein engineering research. We built our advanced Shotgun Mutagenesis technology platform to design protein variants that demonstrate desired properties, such as enhanced expression level or solubility. On this platform, customers can perform comprehensive mutagenesis and high-throughput analysis of any protein of interest, and it is an excellent alternative to time-consuming or expensive protein mapping and engineering applications, and opens discovery pathways for many untapped targets in the proteome.

Profacgen has accumulated lots of experience in scanning point mutations. Our professional technical team can provide customers with high-quality shotgun mutagenesis platform and many related featured services. Our competitive prices and extensive expertise have earned us the trust of our collaborators. Contact us to find out how Profacgen could be of assistance.

References

1. Morrison, K.L.; Weiss, G.A. Combinatorial alanine-scanning. Current Opinion in Chemical Biology. 2001.
2. Davidson, E.; Doranz, B.J. A high-throughput shotgun mutagenesis approach to mapping B-cell antibody epitopes. Immunology. 2014.