Efficient Screening Methods and Research Progress of Anti-tumor Polypeptides-Special Topic on Phage Display Technology-Tekbiotech-Yeast Display Service,Phage display technology

Efficient Screening Methods and Research Progress of Anti-tumor Polypeptides

In recent years, cancer has become younger and younger, and it is one of the diseases with high mortality rates worldwide. The current methods used to treat cancer all have side effects to varying degrees. In comparison, peptide molecules cause mild adverse reactions to the body and have strong targeting, and have become a new type of anti-tumor drug. Anti-tumor peptide drugs have great potential and provide new options for patients with limited or ineffective traditional treatments.

Peptide drugs are usually peptide analogs synthesized from endogenous peptides. They are one of the hot directions in drug development at present, among which anti-tumor peptides have attracted much attention. Anti-tumor peptides have anti-cancer effects and are isolated from a variety of natural sources (such as venom secreted by amphibians and reptiles has anti-tumor activity, and short-tailed hamsters contain peptides that can inhibit the proliferation of ovarian cancer and breast cancer), so they have high selectivity and high penetration efficiency.

Anti-tumor peptides can be divided into inhibitory peptides, necrosis-inducing peptides and apoptosis-promoting peptides. Surgery and chemotherapy are common means of treating diseases. Surgery will cause certain risks, and chemotherapy will cause adverse reactions in the body. Therefore, a new anti-tumor drug, anti-tumor peptides, has emerged. Compared with surgery or chemotherapy, it has similar therapeutic effects, but higher safety and tolerance.

1. Anti-tumor Peptide Extraction and Separation Methods

Currently, the extraction and separation methods of anti-tumor peptides are divided into chemical extraction, enzymatic hydrolysis, ultrafiltration and chromatography according to the physicochemical properties of proteins (charge, isoelectric point, hydrophilicity and molecular weight, etc.). Peptides from nature are often prepared by enzymatic hydrolysis; separation methods are divided into ion exchange, molecular exclusion, reverse phase, affinity chromatography and electrophoresis. Gel filtration and reverse chromatography are often used according to the molecular weight and lipid solubility of peptides.

2. Phage Display Peptide Library Screening

The methods for phage display peptide library screening are as follows:

2.1 In situ screening at the molecular level

In situ screening is to coat a specific target protein on a 96-well plate. This screening method is not easily interfered by non-specific factors because it only contacts a single target, and can obtain peptide ligands with high binding affinity.

2.2 Cell-level screening

Cell-level screening is to screen out peptides that bind to components or surface receptors highly expressed in tumor cells by comparing with normal cells. The peptides obtained by this screening method have targeting and high affinity.

2.3 Overall-level screening

The established phage peptide library is injected into the animal by intravenous injection. After a period of time, the phage is extracted and amplified from the subcutaneous tissue of the animal. After 3-5 rounds of repeated selection, enriched targeting peptides are obtained, completing the process of targeting peptide screening.

3. Peptide Screening Based on Tumor Microenvironment

3.1 Peptide screening based on tumor blood vessels

Taking RGD peptide as an example, RGD peptide can be specifically recognized by integrin and perform regulatory functions in the processes of cell adhesion, invasion, proliferation, survival and apoptosis.

3.2 Peptide screening based on tumor metastasis

Matrix metalloproteinase is one of the upregulated molecules in the tumor microenvironment, which participates in the movement and invasion of tumor cells and can inhibit the invasion of cancer cells by mediating signal pathways.

4. Peptide Screening Based on Biological Phenotype

Compared with traditional phenotypic screening methods, peptide screening technology based on biological phenotype has significant advantages, especially in the field of tumor research and treatment. Traditional phenotypic screening usually relies on the identification of cell surface markers or active molecules, while screening based on biological phenotype pays more attention to changes in cell internal functions and molecular interactions, and can provide more accurate targeted intervention mechanisms.

For tumor treatment, disrupting the interaction between S phase kinase-associated protein 2 (Skp2) and Cyclin A has become a potential therapeutic strategy. Skp2 and Cyclin A play an important role in the S phase of the cell cycle. Skp2, as an important E3 ubiquitin ligase, can promote the cell cycle by regulating the degradation of multiple cell cycle proteins, while Cyclin A interacts with CDK2 to drive cells into the S phase. Therefore, the interaction between these two molecules is a key link in cell cycle regulation.

Studies have shown that by specifically interfering with the binding of Skp2 and Cyclin A, tumor cells can be effectively prevented from entering the S phase, thereby inhibiting their proliferation. Since tumor cells usually have a higher proliferation rate, this intervention can selectively kill tumor cells while having less impact on normal cells, thereby improving the selectivity and safety of treatment. Peptide screening based on biological phenotypes can help discover molecules that specifically interfere with these interactions and provide new therapeutic targets for tumor treatment.

TekBiotech has been committed to the field of antibodies for many years and has rich experience and mature technology. It can provide customers with high-quality peptide library synthesis and screening technology, targeted peptide screening technology, phage display technology, and mRNA display technology services. In addition, TekBiotech has many years of experience in phage cyclic peptide library screening, and can provide peptide library screening services, phage library screening services, and customized antibody services, including antibody expression and purification, affinity determination, antibody sequencing, etc., to meet customer needs.

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