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Targeted Peptide Library Construction Service

Based on its experienced phage surface display technology, TekBiotech provides targeted peptide discovery services to global customers. Different from the well-known recombinant M13 phage display technology, the M13 peptide surface display technology provided by TekBiotech is to modify the wild-type M13 phage and insert the random peptide sequence into the N-terminus of the P3 protein of the wild-type M13 phage through gene editing engineering, so that the random peptide is displayed on the surface of the wild-type M13 phage along with the expression of the coat protein, and then the targeted peptide is screened through a simple blue-white screening system.

TekBiotech has rich project experience and insights in peptide library construction. After years of development, TekBiotech has established a complete peptide library construction system (including but not limited to M13 phage display system, M13 KE phage display system, T7 phage display system, etc.) that relies on M13 helper phage, and can provide customers with high-quality peptide library construction services including but not limited to linear peptide libraries (including but not limited to 6-peptide library, 7-peptide library, 12-peptide library, 15-peptide library) and cyclic peptide libraries (cyclic 6-peptide library, cyclic 7-peptide library, cyclic 10-peptide library, etc.).

The peptide library construction service is a library construction service based on phage display technology, which refers to the use of 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 construct it into a suitable phage vector and transform it into the corresponding host bacteria for packaging and amplification to obtain a phage library. The M13 phage peptide library established by TekBiotech has a library capacity of up to 10^12 and a titer of up to 10^16 phage display peptide particles/ml. The T7 phage peptide library has a library capacity of up to 10^10 and a titer of up to 10^13 phage display peptide particles/ml, which is sufficient to support customers' subsequent screening of targeted peptides for various targets and meet the needs of downstream experiments. TekBiotech 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 TekBiotech scientists will design the best library construction method and phage system according to the customer's project requirements to meet the project needs of different customers.

█ Targeted Peptide Library Construction and Development Service Based On Phage Display Technology

TekBiotech has established two M13 phage display peptide library systems. One is the M13 phage display system that requires M13 auxiliary phage, that is, the synthetic peptide gene library is constructed into the pEMCS vector, and the library construction is completed through transformation, amplification, phage packaging (library rescue), etc. The second is the M13 wild-type phage display system, that is, the designed and synthesized peptide gene library is edited into the M13 phage genome, and the targeted peptide positive clones can be obtained through transformation, amplification and blue-white screening. The construction process of M13 phage display peptide library is shown in Figure 1:

Targeted Peptide Library Construction Service-tekbiotech1.png

Figure1 Construction process of the blue-white class M13 phage display peptide library

█ Service Content and Cycle of the Construction of the Targeted Peptide Phage Library

Classification

Service Content

Cycle

Construction of peptide/cyclic peptide gene mutation library

(1)Trimer codon technology/NNK technology/Error-prone PCR technology for gene library synthesis

(2)Delivery: gene library synthesis quality report, 3ug gene library

5-7 Weeks

Construction of M13 phage peptide/cyclic peptide library

Blue-white class M13 phage display system:

(1) Phage vector construction and transformation

(2) QC identification: monoclonal sequencing + NGS sequencing

(3) Delivery: constructed phage library with target cyclic peptide/linear peptide, experimental report

Recombinant M13 auxiliary phage display system:

(1) Phage vector construction and transformation

(2) Library rescue;

(3) QC identification: monoclonal sequencing + NGS sequencing

(4) Delivery: constructed phage library with target cyclic peptide/linear peptide, experimental report

3-6 Weeks

█ Targeted peptide discovery technology platform service advantages

Customization of various types of peptide

libraries: construction of linear peptide

libraries and cyclic peptide libraries

High quality of gene libraries: chip method

and Trimer Codon synthesis

Optional display systems: M13 phage

system (wild-type M13 system,

recombinant phage system), T7 phage

system

One-to-one personalized solution

customization to meet the scientific

research project needs of various

customers

Targeted Peptide Library Construction Service Frequently Asked Questions

What is Targeted Peptide Library Construction Service? What is its main application?

Targeted peptide library construction service is to screen out peptide molecules with high affinity and specificity to specific targets by synthesizing libraries containing a large number of peptide sequences. This service is usually applied in the fields of antibody development, targeted therapy, vaccine research, and disease marker discovery. In antibody development, targeted peptide libraries are used to screen peptides that bind specifically to antigens to support the discovery of antibody drugs; in targeted therapies, peptides can be used as a drug delivery system to deliver therapeutic molecules to target cells with precision; and in vaccine research, peptides can be used as antigens to help identify potential immune targets.
What is the core technology of targeted peptide library construction service?

The construction of targeted peptide libraries relies on high-throughput peptide synthesis technology, usually using solid-phase synthesis or liquid-phase synthesis. In solid-phase synthesis, the peptide chain is synthesized step by step using chemical reactions, and the amino acid residues are added to the solid-phase carrier one by one. In contrast, liquid-phase synthesis realizes peptide synthesis through chemical reactions in solution. Library construction also includes a randomization process, whereby a diverse library of peptides is generated by varying the synthesized amino acid sequences to ensure coverage of a wide range of structural features. During the library construction process, the synthesized peptides are usually expressed and screened using phage display technology or yeast display technology to identify high-affinity peptides that bind to specific targets.
How to select suitable targets for screening of targeted peptide libraries?

Target selection is the most critical step in the construction and screening of targeted peptide libraries. The selection of target needs to take into account its role in the disease, surface-exposed region and biological function. Common targets include tumor-associated antigens, bacterial toxins, viral surface proteins and so on. When selecting targets, firstly, it is necessary to ensure that they are specific and able to distinguish between target cells and normal cells; secondly, targets should have enough surface-exposed area for peptides to bind to them. The solubility and stability of the target should also be taken into consideration, which will help efficient binding and expression in the subsequent screening process. By selecting appropriate targets, the success rate of screening high affinity and multifunctional targeted peptides can be improved.
How can targeted peptide libraries ensure high-affinity and specific screening results?

To ensure the high affinity and specificity of targeted peptide libraries, first of all, at the library construction stage, a large number of amino acid changes will be introduced during the synthesis of peptides to ensure the diversity of the libraries. During the screening process, techniques such as reverse elution are used to improve the specificity of the screening, thus ensuring that the screened peptide can have strong binding to the target. To address the problem of non-specific binding, an elution step can be introduced in the screening process to remove peptides that bind to non-target substances, ensuring the purity of the library. In addition, after screening, the peptides need to be subjected to affinity assay to ensure that their binding constants (Kd) to the target are low enough to obtain targeted peptides with high affinity and specificity.
How to evaluate the function and effect of the screened targeted peptide?

Functionality and efficacy of screened target peptides are usually evaluated through a series of biological experiments. First, the binding affinity of the peptide to the target is detected by ELISA, surface plasmon resonance (SPR), fluorescence resonance energy transfer (FRET) and other methods. Second, the biological activity of the peptide is assessed by cellular assays, such as by flow cytometry, immunofluorescence staining, etc., to observe whether the peptide specifically recognizes and binds to the target cells or cell surface markers. The in vivo stability and pharmacokinetic properties of peptides can also be assessed by animal models. Through these experiments, the effects and potential applications of the targeted peptides can be comprehensively evaluated.

What is Targeted Peptide Library Construction Service? What is its main application?

Targeted peptide library construction service is to screen out peptide molecules with high affinity and specificity to specific targets by synthesizing libraries containing a large number of peptide sequences. This service is usually applied in the fields of antibody development, targeted therapy, vaccine research, and disease marker discovery. In antibody development, targeted peptide libraries are used to screen peptides that bind specifically to antigens to support the discovery of antibody drugs; in targeted therapies, peptides can be used as a drug delivery system to deliver therapeutic molecules to target cells with precision; and in vaccine research, peptides can be used as antigens to help identify potential immune targets.
What is the core technology of targeted peptide library construction service?

The construction of targeted peptide libraries relies on high-throughput peptide synthesis technology, usually using solid-phase synthesis or liquid-phase synthesis. In solid-phase synthesis, the peptide chain is synthesized step by step using chemical reactions, and the amino acid residues are added to the solid-phase carrier one by one. In contrast, liquid-phase synthesis realizes peptide synthesis through chemical reactions in solution. Library construction also includes a randomization process, whereby a diverse library of peptides is generated by varying the synthesized amino acid sequences to ensure coverage of a wide range of structural features. During the library construction process, the synthesized peptides are usually expressed and screened using phage display technology or yeast display technology to identify high-affinity peptides that bind to specific targets.
How to select suitable targets for screening of targeted peptide libraries?

Target selection is the most critical step in the construction and screening of targeted peptide libraries. The selection of target needs to take into account its role in the disease, surface-exposed region and biological function. Common targets include tumor-associated antigens, bacterial toxins, viral surface proteins and so on. When selecting targets, firstly, it is necessary to ensure that they are specific and able to distinguish between target cells and normal cells; secondly, targets should have enough surface-exposed area for peptides to bind to them. The solubility and stability of the target should also be taken into consideration, which will help efficient binding and expression in the subsequent screening process. By selecting appropriate targets, the success rate of screening high affinity and multifunctional targeted peptides can be improved.
How can targeted peptide libraries ensure high-affinity and specific screening results?

To ensure the high affinity and specificity of targeted peptide libraries, first of all, at the library construction stage, a large number of amino acid changes will be introduced during the synthesis of peptides to ensure the diversity of the libraries. During the screening process, techniques such as reverse elution are used to improve the specificity of the screening, thus ensuring that the screened peptide can have strong binding to the target. To address the problem of non-specific binding, an elution step can be introduced in the screening process to remove peptides that bind to non-target substances, ensuring the purity of the library. In addition, after screening, the peptides need to be subjected to affinity assay to ensure that their binding constants (Kd) to the target are low enough to obtain targeted peptides with high affinity and specificity.
How to evaluate the function and effect of the screened targeted peptide?

Functionality and efficacy of screened target peptides are usually evaluated through a series of biological experiments. First, the binding affinity of the peptide to the target is detected by ELISA, surface plasmon resonance (SPR), fluorescence resonance energy transfer (FRET) and other methods. Second, the biological activity of the peptide is assessed by cellular assays, such as by flow cytometry, immunofluorescence staining, etc., to observe whether the peptide specifically recognizes and binds to the target cells or cell surface markers. The in vivo stability and pharmacokinetic properties of peptides can also be assessed by animal models. Through these experiments, the effects and potential applications of the targeted peptides can be comprehensively evaluated.

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