As an undisputed forerunner in phage display and antibody manufacturing, Creative Biolabs has been consistently dedicated to exploring groundbreaking scientific advances while providing comprehensive services. Our scientists are now proud to offer high-class customized service of T7 phage display library construction to support diverse research projects of our worldwide clients.
Phage display is a powerful laboratory approach for identifying multiple molecules that have desirable binding properties. Despite filamentous phages (M13, fd, f1) have long been the most frequently used vehicle, display systems based on other types of phages, especially bacteriophage T7, are gaining more and more attention for their unique features.
Similar to T4, bacteriophage T7 possesses a head and tail structure. The icosahedral head, where T7 conserves its dsDNA (39937 bp) genome, is composed of 415 copies of capsid gp10, arranged as 60 hexamers on the surface and 11 pentamers at the vertices. Tails are shorter, and not contractible, hence a protein channel from tail to the host cytoplasm would be built during infection. There exist two isoforms of major coat protein gp10, gp10A and gp10B, resulted from natural translational frameshift at amino acid 341. However, functional capsid can be made from either 10A or 10B or complex of random ratios, implying that T7 capsid shell can accommodate variations. Besides, T7 undergoes a lytic lifecycle, and is extremely robust, which can form plaques within 3 h at 37 °C and the release progeny phages 1-2 h after infection, thus significantly reducing the time required for selection. It can also endure harsh conditions that inactive other phages.
T7 phage display system involves the capsid isoform gp10B to display foreign proteins or peptides. The natural translational frameshift site of the gp10 gene is removed in the vector sequence to ensure the only gp10B is expressed. Creative Biolabs offers multiple vector choices for display of different insertion sizes, with different display densities and host bacteria strains. After preparation of recombinant vectors, our scientists use appropriative in vitro packaging extracts to enable efficient construction of high-capacity libraries. The addition of special tags helps to verify the library diversity and achieve high-throughput screening. We also conduct specific biopanning protocols to isolate target antibodies or peptides with ideal properties required by customers.
Major advantages of T7 phage display system for library construction including but not limited to:
According to the advanced technical platforms and extensive expertise in phage display field, Creative Biolabs is the right company who can provide high quality and goal-oriented services to boost your library construction projects. Scientists in Creative Biolabs are looking forward to sharing our professional knowledge and providing one-stop solutions for our global customers.
Fig. 1 Classifiers to predict TB from healthy controls and sarcoidosis patients.1
The research aimed to identify biomarkers for active tuberculosis (TB) using a novel T7 phage display cDNA library derived from sarcoidosis and Mycobacterium tuberculosis (MTB) components. The study successfully identified specific antigenic clones that could distinguish active TB patients from sarcoidosis patients and healthy controls with high sensitivity and specificity. The T7 phage display library enabled the construction of a complex sarcoidosis library (CSL) and the identification of unique antigens from bronchoalveolar cells and white blood cells. These antigens were used to create a microarray platform for immunoscreening, leading to the discovery of potential diagnostic biomarkers for TB. The T7 phage display library enhances the detection and differentiation of TB, offering a promising tool for improving TB diagnosis and potentially guiding the development of targeted therapies and vaccines.
A T7 phage display library is a powerful molecular tool used to identify peptide or protein interactions with specific targets. It involves inserting a cDNA or peptide-encoding sequence into the genome of the T7 bacteriophage, which then displays the encoded peptide on its surface. This allows researchers to screen and select peptides or proteins that have a high affinity for a particular target, facilitating the discovery of novel binding partners, therapeutic candidates, or diagnostic markers.
The T7 phage display library is widely used in drug discovery, vaccine development, and biomarker identification. It is particularly valuable for identifying peptides or proteins that bind to specific molecular targets, such as antigens in pathogens or disease-related proteins. This technology is also employed in the development of diagnostic assays and the study of protein-protein interactions, making it a versatile tool in both basic and applied sciences.
Construction of a T7 phage display library involves inserting DNA sequences encoding peptides or proteins of interest into the genome of the T7 bacteriophage. These sequences are inserted into specific sites in the phage genome, allowing the encoded peptides to be displayed on the surface of the phage. The library is then amplified in Escherichia coli, resulting in a diverse collection of phages, each displaying a different peptide. This library can be screened to identify phages that bind to a specific target.
The T7 phage display library offers rapid amplification and high-density display of peptides, which enhances the efficiency of binding interactions. The T7 phage has a robust replication cycle and can produce large quantities of phage particles in a short time, making it suitable for high-throughput screening. Additionally, the T7 system is less prone to contamination by other bacterial phages, which can interfere with the selection process, providing a cleaner and more reliable platform for peptide selection.
The T7 phage display library allows researchers to screen vast numbers of peptides or proteins against a specific target, such as a disease-related protein. By selecting peptides that bind strongly to the target, researchers can identify potential therapeutic candidates that could inhibit or modulate the target's function. This approach is particularly valuable in drug discovery, where the identification of novel binding partners can lead to the development of new drugs or therapies.
One of the main challenges is ensuring the proper folding and functionality of the displayed peptides, as incorrect folding can lead to false-negative results. Additionally, the complexity of the target protein or the biological environment in which the screening is conducted can affect the selection process. Careful optimization of screening conditions and validation of selected peptides are necessary to overcome these challenges.
In vaccine development, the T7 phage display library is used to identify peptide epitopes that can elicit a strong immune response. Researchers screen the library against antigens from pathogens to select peptides that mimic these antigens and can induce protective immunity. These selected peptides can then be incorporated into vaccine formulations, providing a targeted and effective approach to vaccine design. This method is particularly useful for developing vaccines against pathogens with complex antigenic profiles.
Biopanning can enrich for phages that display peptides with high affinity for a target. The process involves incubating the phage library with the target of interest, washing away non-binding phages, and then eluting the bound phages. These eluted phages are amplified and subjected to additional rounds of selection to further enrich for high-affinity binders.
By screening the T7 phage display library against sera or tissue samples from patients, researchers can select peptides that bind specifically to disease-related antigens. These selected peptides can then be used to develop diagnostic assays that detect the presence of the disease, improving early diagnosis and treatment.
The T7 phage display library has the potential to be applied in emerging areas of research, such as personalized medicine, where it can be used to identify patient-specific therapeutic targets or biomarkers. Additionally, the integration of T7 phage display with high-throughput sequencing and artificial intelligence could accelerate the discovery of novel peptides and proteins with therapeutic or diagnostic potential. Future developments may also include the use of T7 phage display in synthetic biology, where precise control over peptide sequences is essential for engineering new biological functions.
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