Precise Mutations at Multiple Sites with Various Amino Acids:

Simultaneously mutate several consecutive amino acid sites within a defined region of a protein sequence into any of the other 19 amino acids. This process forms a combinatorial mutation library, having multiple amino acid sites mutated simultaneously. This causes each sequence to exhibit multiple amino acid site mutations when compared to the template.

This provides a powerful experimental tool for studying the relationship between complex protein structure and function and for research in protein engineering and antibody such as the Trimer library.

Directed Evolution Library of AAV Capsid Protein

Synbio Technologies offers a comprehensive platform for the design and synthesis of variant libraries. We provide customized, one-stop services for designing, constructing, validating, viral packaging, and bioinformatics analysis of directed evolution libraries specifically for AAV capsid proteins across 12 different AAV serotypes (AAV1-AAV12). With our platform, customers can easily screen for novel and superior AAV capsids, such as those with tissue specificity, immune evasion, and transgene expression, thereby accelerating the development of better AAVs. In addition to our custom services, we have launched a ready-to-use peptide display library for AAV9 capsid proteins. This library is an invaluable tool for optimizing AAV gene delivery vectors, making it easier for researchers to achieve more efficient gene delivery solutions.

 AAV9 Capsid Protein Peptide Display Library

Case Study

Case Study 1

The NNK library is a classic degenerate mutation library. The NNK degenerate primers used contain 32 (4x4x2) codon combinations (N= A/C/G/T, K= G/T), covering all 20 amino acids with the ability to saturate amino acids at any site.

Fig.1 The proportion of each codon contained in the five NNK sites in the NNK library

Case Study 2

Trimer libraries can be used for site saturation mutation to distribute 20 amino acids in equal proportion, further reducing codon redundancy and removing termination codons in the library.

 Fig.2 Trimer libraries are also a saturated mutation library with equal distribution of each amino acid

Case Study 3

Trimer libraries can also customize each site to be composed of different ratios of amino acids. Among the four Trimer sites, for example, the X1 Trimer site is A (80%) : G (3%) : R (3%) : S (3%): T (8%) : V (3%)

Fig.3 Types and proportions of custom amino acids

 Fig.4 Ratio between actual and theoretical proportion of each amino acid

FAQs

What is a precision variant libraries?

Precision variant libraries are a large collection of mutated homologous genes constructed through modern molecular biology methods. These libraries simulate natural evolution mechanisms by introducing mutations at specific sites or regions; they are often used to study the structure-function relationships of proteins or other biomolecules, providing important experimental tools for fields such as enzyme engineering, biomedicine, agriculture, and food science.

How to choose suitable precision variant libraries?

Selecting the suitable precision variant library depends on specific research objectives and experimental needs. For instance, if the goal is to explore the role of a single amino acid in protein function, an alanine scanning library might be the ideal choice. However, if the aim is to study the function of multiple amino acid sites, a site-saturation mutagenesis library or a combinatorial mutation library would be appropriate. Additionally, other factors must be considered, such as the availability of the mutation library, the robustness of the screening method, and the experimental costs. By carefully evaluating these elements, researchers can choose the most suitable variant library to achieve their scientific objectives effectively.

What are the applications of precision mutation libraries?

Some applications of precision mutation libraries are as follows:
 • Protein Engineering: used to improve the catalytic properties, stability, and specificity of enzymes.
 • Drug Development: used to screen the key functional areas of drug targets and discovering new drug targets related to diseases.
 • Antibody Engineering: helps construct high affinity and specificity antibody libraries for immunotherapy or diagnosis.
 • Basic Scientific Research: helps explore the function, regulatory elements, and regulation mechanism of genes to aid the progress of basic scientific research.