M13 Phage Display Peptide Library Screening Platform
Alpha Lifetech brings over a decade of experience in phage high throughput library screening.
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M13 Phage Display Peptide Library Screening Platform
There are many significant advantages of Alpha Lifetech in life science research and biomedical development due to its flexibility, reliability and efficiency. Tailored specific project requirements can also be arranged based on customer demands.
M13 phage display technology is an efficient molecular screening platform based on M13 phage, which is widely used in biology, drug development and materials science. By using phage display peptide library, Alpha Lifetech can achieve high specific binding of target molecules to specific peptides, so as to screen out peptide molecules with excellent functions. The core of the M13 phage display technology is to realize the directed evolution between molecules by combining the random peptide library with the target ligand.
The core of m13 phage display library
M13 phage is a filamentous phage, and its unique structure and biological characteristics make it an ideal carrier for peptide display. In the M13 phage display system, random peptide sequences are inserted into the gene sequence of the outer membrane protein ( usually pIII or pVIII ) of the phage to display on the surface of the phage. These displayed peptides form a huge phage display peptide library, which contains millions or even billions of different peptide sequences.The species display method directly links the genotype ( DNA sequence encoding the peptide ) to the phenotype ( structure and function of the peptide ), making the screening process efficient and accurate. By combining high throughput screening technology, M13 phage display technology can identify high-affinity binding peptides of target molecules from a large peptide library in a short time.
Fig.1 Schematic showing the infection and production process of natural M13 phage. Illustration of the mechanism of phage vector-based display system. Illustration of the mechanism of phagemid-based display system. ( Reference source: Screening Phage-Displayed Combinatorial Peptide Libraries. )
Construction and screening process of phage peptide library
Random peptide sequence design
DNA encoding diverse peptides was generated by synthesizing random nucleotide sequences. Typically, these peptides are 7-12 amino acids long.
Insertion of phage vector
The random peptide sequence was inserted into the outer membrane protein gene of M13 phage so that it could be displayed on the phage surface. A typical phage peptide library contains about 10^8 to 10^10 different peptide sequences to ensure that the target molecule can find specific binding peptides.
Target fixation
The target molecules ( such as proteins, receptors or antibodies ) were immobilized on solid-phase carriers ( such as magnetic beads, microplates or nylon membranes ). The form of fixed targets can enhance the specificity of screening.
The process of incubating the phage display peptide library
The constructed phage display peptide library was incubated with the fixed target, allowing the peptides on the phage to bind to the target molecule. In this process, the unbound phages were removed by elution, while the phages bound to the target were retained.
Elution and recovery
High affinity binding phages were recovered using specific elution conditions ( such as pH gradient, competitive inhibitors or denaturants ).
Amplification and rescreening
The recovered phages were amplified by infecting E.coli, followed by the next round of screening. After each round of screening, the retained phage population has higher specificity and affinity. Usually, 3-5 rounds of screening are required to obtain peptides with high binding efficiency.
Sequencing and functional verification
After the peptide library screening is completed, in order to analyze the results in depth, high-throughput screening methods, especially high-throughput sequencing techniques, are usually used to comprehensively analyze the screened phage libraries. High-throughput sequencing of selected phages is usually performed. High-throughput sequencing can quickly and accurately detect phage sequences enriched in phage peptide libraries, thereby revealing peptide sequences or antibody fragments that are highly bound to the target. In addition, by analyzing the frequency distribution and enrichment trend of sequences, the efficiency of the screening process can be evaluated, and the function and target affinity of specific sequences can be further verified. By combining peptide library screening and high-throughput screening, researchers can efficiently mine functional peptide sequences from complex phage peptide libraries, laying a foundation for drug development, biomarker discovery and functional research.
Fig.2 Scheme of the SCOPE system-based phage screening for micropillar array and particle tracing. ( Reference source “M13 phage: a versatile building block for a highly specific analysis platform. )
High Cost-Effectiveness
Compared with other technologies ( such as antibody preparation ), M13 phage display technology of Alpha Lifetech has a shorter development cycle and lower overall cost, which is especially suitable for preliminary screening and functional verification.
Mature Analysis System
After screening, the abundance and distribution of the selected peptides can be analyzed by next-generation sequencing ( NGS ) to quickly lock high-affinity peptides.
The platform also provides subsequent verification methods, such as ELISA and SPR, to ensure the reliability of the screening results.
High Sensitivity and Specificity
Using the high stability and natural amplification ability of phage particles, even if the target molecule concentration is low, the target binding peptide can still be detected and screened by phage amplification technology.
Diversity Target Molecules
The platform is suitable for a variety of target molecules, such as proteins, small molecules, nucleic acids and even complex cell surface receptors.
It can screen functional peptides suitable for different targets and is widely used in drug development, diagnostic reagent development and biomarker research.
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2018-07-16
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