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Scanning Point Mutation Libraries

Creative Biostructure has developed protein evolution and engineering platform through many years of experience, our seasoned scientists can provide tailored mutagenesis library construction services based on scanning point mutation technique.

Scanning point mutation is a excellent and systematic means of improving protein properties, which can replace each amino acid of target protein with all 20 amino acids simultaneously. This powerful technique can offer a detailed profile of each amino acid at the position. For each desired codon, a small, site-saturated library can be constructed. Scanning point mutation library can be delivered by Creative Biostructure as a pool or in a separated format for any substitution variant. Creative Biostructure can provide superb sequential permutation scanning library construction services.

Notably, Creative Biostructure has strong expertise in custom alanine (Ala) scanning services for protein engineering. Ala scanning technique is popularly used high-throughput mutagenesis method in which amino acid residues in a target protein are systematically replaced for alanine at selected positions by site-directed mutagenesis, expressed, and analyzed for protein function. It is reported that substitution with alanine residues can eliminate side-chain (R = methyl) interactions without changing main-chain conformation or introducing electrostatic or steric effects, therefore, which is often the preferred choice for determining the contribution of specific side-chains while retaining native and entire protein structure. Alanine scanning can be used for testing activation and coupling, interactions with ligands and monoclonal antibodies (mAbs).

Scanning Point Mutation Libraries Figure 1. Alanine scanning used in phage display.

Many structurally complex proteins are thousands of amino acids amino acids long, individual alanine substitutions across their entire length will be laborious and time-consuming, typically limiting functional analysis of target proteins. Thus, Creative Biostructure can use combinatorial libraries of mutant proteins to rapidly analyze protein function and identify important sidechain functionalities. Combinatorial libraries of alanine substitutions are an alternative to the laborious method of scanning individual positions in a protein. There are two approaches to perform combinatorial libraries: binomial mutagenesis and shotgun scanning. Shotgun scanning is a simplified method to identify epitope and functional mapping. Creative Biostructure ensures that using shotgun mutagenesis to engineer protein variants for improving desired properties such as enhanced expression or solubility.

Creative Biostructure also provides Membrane Protein Platform and Phage Display Platform for your specific projects. Please feel free to contact us for a detailed quote.

References:
E. Berdougo and B. J. Doranz (2012). High-Throughput Alanine Scanning. Genetic Engineering & Biotechnology New. Vol. 32, No. 12
K. L. Morrison and G.A. Weiss (2001). Combinatorial alanine-scanning. Curr. Opin. Chem. Biol., 5: 302-307.
L. Q. Al-Mawsawi, et al. (2014). High-throughput profiling of point mutations across the HIV-1 genome. Retrovirology, 11: 124. DOI 10.1186/s12977-014-0124-6


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