Phage Display Peptide Library Construction

* Please kindly note that our products and services can only be used to support research purposes (Not for clinical use).

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Creative Peptides is committed to providing customers with high-quality peptide library construction technical services and providing strong support for customers' subsequent downstream research and development work such as peptide library screening, targeted peptide drug discovery, specific blocking peptide drug development, and specific lead sequence development. Creative Peptides has rich project experience and experience in peptide library construction. After years of development, Creative Peptides has established a complete peptide library construction system (including M13 phage display system, M13 KE phage display system, T7 phage display system), can provide customers with high-quality linear peptide libraries (including 6-peptide libraries, 7-peptide libraries, 12-peptide libraries, 15-peptide libraries) and cyclic peptide libraries (cyclic 6-peptide library, cyclic 7 peptide library, cyclic 10 peptide library) and other types of peptide library construction services.

The phage display peptide library construction service refers to using gene mutation methods such as Trimer Codon technology, NNK technology or Error-prone PCR technology to synthesize a gene library that meets customer needs, and then constructs it into a suitable phage vector and Phage library obtained by transforming into corresponding host bacteria for packaging and amplification. The M13 phage peptide library established by us has a library capacity of up to 10⁸ and a titer of 10¹³ phage display peptide particles/ml. The T7 phage peptide library has a library capacity of up to 10⁸ and a titer of 10¹¹ phages. The display of peptide particles/ml is sufficient to support customers' subsequent screening of targeted peptides for various targets and to meet the needs of downstream experiments. Creative Peptides can provide one-stop technical services including peptide gene library design and synthesis, peptide library construction, supporting peptide library screening, affinity verification, in vitro cell verification, etc. Customers only need to provide specific project requirements, and Creative Peptides scientists will design the best library construction method and phage system according to customers' project needs to meet the project needs of different customers.

Phage display peptide library construction protocol

Our phage display peptide library construction protocol outlines the step-by-step process we follow to construct high-quality phage display libraries for our clients. This protocol has been refined and optimized based on years of experience and expertise in the field of phage display technology.

• Design and selection of library format

Project consultation: We begin by consulting with the client to understand their project goals, target molecules, and desired library format (e.g., linear peptides, cyclic peptides, target-specific peptides).

Library design: Based on the project requirements, our team designs the library format, including the length and diversity of peptide sequences, linker sequences and any specific motifs or modifications required.

• Gene synthesis and library construction

Gene synthesis: Synthetic genes encoding the designed peptide sequences are synthesized using state-of-the-art DNA synthesis technology.

Library assembly: The synthetic genes are then assembled into a phage display vector using molecular cloning techniques. The vector contains the necessary elements for phage display, including a phage coat protein fusion site for peptide display.

• Transformation and phage packaging

Transformation: The constructed phage display vector is transformed into a suitable Escherichia coli (E. coli) host strain for phage production.

Phage packaging: The transformed cells are infected with helper phage to initiate phage production. The helper phage provides the necessary components for packaging the phage display vector into infectious phage particles.

• Library amplification

Phage amplification: The infected E. coli cells are cultured to allow for the amplification of the phage display library. During this process, the phage particles display the peptide sequences encoded by the synthetic genes on their surfaces.

• Library characterization and quality control

Library titration: The titer of the phage display library is determined by plaque assay or other quantitative methods to assess library diversity and complexity.

Sequencing: A subset of phage clones may be sequenced to verify the diversity and integrity of the library.

Functional validation: The functionality of the phage display library may be validated through binding assays using known targets or control ligands.

• Delivery and client support

Library delivery: The constructed phage display library is delivered to the client along with a detailed report summarizing the construction process, quality control data, and any additional information relevant to the project.

Client support: Our team provides ongoing support to the client, addressing any questions or concerns that may arise during the use of the phage display library.

Our phage display peptide library construction

At Creative Peptides, we offer comprehensive phage display peptide library construction services. Our capabilities include creating various types of peptide libraries, customized to meet the specific needs of each project. We are proficient in tailoring the most suitable phage display system for library construction, including M13, T4, T7, or lambda phages.

Types of phage display peptide libraries

Linear peptide libraries: These libraries consist of short, linear peptide sequences displayed on the surface of phages. Linear peptide libraries are essential for mapping epitopes, identifying enzyme substrates, and discovering peptide ligands.

Cyclic peptide libraries: These libraries display cyclic peptides, which are formed by introducing a covalent bond between the N and C-termini or side chains of the peptide. Cyclic peptide libraries are valuable for identifying peptides with enhanced stability and affinity.

Random peptide libraries: These libraries contain randomly generated peptide sequences, providing a broad diversity that is ideal for screening a wide range of potential interactions.

Target-specific peptide libraries: These libraries are designed with peptides that have a known or predicted affinity for a specific target, facilitating the rapid identification of high-affinity binders.

Library capacity and affinity

Our phage display peptide libraries boast a large capacity, ranging from 10⁸ to over 10¹³ unique clones. This extensive diversity ensures that even rare binders can be identified. Additionally, our libraries achieve high affinity, with dissociation constants (Kd) typically ranging from 10^-7 to 10^-9, ensuring the identification of strong and specific interactions.

Peptide/cyclic peptide gene mutation library construction

We use Trimer codon technology/NNK technology/Error-prone PCR technology for gene library synthesis, and deliver gene library synthesis quality reports, 3ug gene library and other data.

M13 phage peptide/loop peptide library construction

M13 KE phage display system

• Phage vector construction and transformation
• QC identification: single clone sequencing + NGS sequencing
• Delivery: Constructed phage library with target cyclic polypeptide/linear polypeptide, experimental report

Display system that requires M13 helper phage

• Phage vector construction and transformation
• Library rescue;
• QC identification: single clone sequencing + NGS sequencing
• Delivery: Constructed phage library with target cyclic polypeptide/linear polypeptide, experimental report

Why choose Creative Peptides?

• A variety of peptide libraries can be customized.
• A variety of mutation systems are available: Trimer Codon technology continuous saturation mutation and non-continuous saturation mutation, NNK technology saturation mutation, Error-prone PCR technology mutation to construct different mutation peptide libraries to meet the needs of different customers.
• Various phage systems: M13 phage system (wild-type M13 phage system, auxiliary phage system), T7 phage system.
• A variety of vectors are available: pEMCS vector (requires helper phage system), M13 KE vector system.
• High-quality library: M13 phage library has a library capacity of up to 10⁸, a titer of up to 10¹³ phages/ml, and library diversity >90%; T7 phage library has a library capacity of up to 10⁸, and a titer of up to 10¹¹ phage/ml, library diversity >90%.
• Supporting downstream screening and verification experiments: peptide library screening (solid phase screening, liquid phase screening, cell screening, animal in vivo screening), affinity verification experiments.
• One-to-one personalized solution customization (including peptide length and structure design, gene mutation solution design, library construction and screening solution design, and downstream affinity verification, blocking verification solution design) to meet the scientific research projects of various customers need.

FAQ

1. What is phage display peptide library construction?

Phage display peptide library construction involves creating a collection of peptides displayed on the surface of bacteriophages (viruses that infect bacteria). This technology allows the identification of peptides that bind to specific targets, facilitating drug discovery, vaccine development, and protein-protein interaction studies.

2. What types of libraries can you construct?

We can construct various types of libraries, including random peptide libraries and custom peptide libraries tailored to your specific needs. Each library type serves different purposes and can be optimized for specific applications.

3. What are the typical applications of phage display peptide libraries?

• Identification of high-affinity ligands
• Epitope mapping
• Antibody development
• Protein-protein interaction studies
• Target discovery for drug development
• Vaccine research

4. What input materials do you require from customers?

Typically, we require the sequence information for the peptides or the target protein for which the library is being constructed. Additionally, if a custom library is needed, any specific requirements or preferences should be communicated at the outset.

5. What is the turnaround time for constructing a phage display peptide library?

The turnaround time can vary depending on the complexity of the library and the specific requirements of your project. Generally, it takes about 4-8 weeks from the receipt of the necessary materials to the completion of the library construction.

6. How do you ensure the quality and diversity of the constructed libraries?

We use advanced techniques and rigorous quality control measures to ensure high diversity and quality of our libraries. This includes deep sequencing, which allows us to verify the presence of a wide variety of unique peptides within the library.

7. Can you provide assistance with downstream applications, such as screening and target identification?

Yes, we offer comprehensive support services that include library screening, target identification, and characterization of binding interactions. Our team of experts can guide you through each step of the process to ensure successful outcomes.

8. What information do you include in the final deliverable?

The final deliverable includes a detailed report with information on the construction process, quality control results, diversity analysis, and any relevant data from initial screenings. Additionally, we provide the actual phage library in the format requested.