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Mirror-Image Phage Display Service

Introduction Service Workflow Applications Troubleshooting & Tips Popular Services Published Data FAQs Resources Related Sections

Phage display is a technology with a long and successful history that has become a staple of molecular biology and drug discovery. It can identify peptides, antibodies, and other ligands that bind to a target of interest. While phage display has long been an essential tool in therapeutic development, Mirror-image phage display (MIPD) is a newer advancement with an advantage over traditional phage display for finding high-affinity ligands, particularly to create D-peptide therapeutics. We're excited here at Creative Biolabs about the potential for this technology in early-stage research. We would love to show you how our mirror-image phage display platform can speed your project up by generating the stable, functional D-peptide tools you need to hit that next all-important milestone. Alongside our specialized mirror-image phage display platform, we offer a comprehensive suite of custom phage display services to meet your research needs.

What is Mirror-Image Phage Display?

Mirror-image phage display is an advanced version of the traditional phage display that uses D-peptides (mirror-image peptides) to identify high-affinity ligands. This technology has several advantages over conventional phage display, resulting in peptides that are more resistant to proteolysis and immune system recognition. Mirror-image targets allow peptides to be more stable and durable in therapeutic and diagnostic applications. By taking advantage of these characteristics, MIPD enables the discovery of peptides with increased specificity, reduced non-specific binding, and stability in harsh environments. MIPD is commonly used in developing peptide therapeutics, antibody generation, cancer-targeting, immunotherapy, and diagnostic assays, where these qualities are particularly valuable. By overcoming some of the limitations of L-peptides, mirror-image phage display provides a powerful tool for discovering novel ligands and optimizing them for clinical use.

Fig.1 Flowchart of the mirror-image phage display mechanism. (OA Literature)Fig.1 The mechanism of mirror-image phage display for D-peptide discovery.1,3

Our Mirror-Image Phage Display Service

Creative Biolabs provides cutting-edge mirror-image phage display services tailored to your research needs. Whether you are looking for peptide discovery, antibody development, or targeted therapeutic applications, our service can provide you with the high-quality, high-affinity ligands essential for your project. Our service includes:

  • Custom Phage Display Library Construction: We design and construct large D-peptide libraries displayed on phages. These libraries can be customized for your specific target, ensuring access to a broad range of potential ligands.
  • Our state-of-the-art screening platforms help identify high-affinity binders from complex peptide libraries.
  • We provide accurate binding affinity data to ensure that your selected peptides or antibodies have the desired specificity and stability.
  • Once you identify potential candidates, we offer optimization services to enhance their binding properties, stability, and specificity.

How Mirror-Image Phage Display Works

We use an indirect approach that lets us use a standard, powerful screening tool (phage display) to end up with the perfect right-handed peptide. It's a four-step process rooted in the beautiful symmetry of biochemistry.

Step 1: Synthesis of the Mirror-Image Target

Everything starts with your target—the disease-related protein you want to inhibit. It's a natural protein made of left-handed amino acids. Our expert chemists get to work and build a perfect, synthetic mirror image of it, using only right-handed D-amino acids. This right-handed target becomes the bait for our screen.

Step 2: Screening the L-Peptide Library Against the D-Target

With the right-handed target ready, we use a massive library of billions of left-handed peptides displayed on viruses called phages. We mix this library with our right-handed target. Through biopanning, we wash away all the peptides that don't stick and keep only the ones that bind strongly.

Step 3: Identification and Sequencing of L-Peptide Hits

After a few rounds of this "fishing," we isolate the best binding phage. We then read its genetic code to find the exact sequence of the winning left-handed peptide. This sequence is our blueprint.

Step 4: Get the Final D-Peptide Candidate

This is the magic step. We take the sequence from the winning left-handed peptide and chemically build a brand-new peptide made entirely of right-handed D-amino acids.

Applications of Mirror-Image Phage Display

Mirror-image phage display (MIPD) has opened new avenues in peptide and antibody discovery, especially for applications where stability, specificity, and resistance to degradation are essential. Some of the significant applications of MIPD include:

  • Peptide therapeutics development
  • Cancer targeting and immunotherapy
  • Antibody development
  • Diagnostic tools
  • Protein engineering

Troubleshooting and Optimization Tips

While mirror-image phage display is a powerful method for discovering high-affinity peptides and antibodies, several challenges may arise during its application. Below are some common troubleshooting tips and strategies for optimizing your experiments:

Problem Possible Causes Solutions
Low Library Diversity Insufficient variety in peptide sequences
  • Increase library size with more diverse peptide sequences.
  • Consider using different peptide scaffolds.
Low Phage Yield Inefficient infection or improper handling of phage libraries
  • Optimize transformation conditions.
  • Ensure proper bacterial culture and phage propagation.
Non-Specific Binding Weak binding or irrelevant peptides isolated
  • Increase washing stringency during screening.
  • Use competitive inhibitors to block non-specific interactions.
Poor Binding Affinity Inadequate library or insufficient optimization
  • Perform multiple rounds of selection to enrich high-affinity binders.
  • Increase stringency for subsequent selections.
Target Instability Target protein degradation during the process
  • Stabilize target with appropriate buffers or stabilizing agents.
  • Ensure proper handling of the target protein.
Optimization of Selected Peptides Peptides require fine-tuning for further use
  • Use techniques like truncation, substitution, or cyclization.
  • Perform further affinity and stability testing.

Popular Services You May Need

At Creative Biolabs, we offer a wide range of related services that complement our mirror-image phage display service, enabling you to develop your peptide and antibody candidates fully.

Service Description
Phage Display Library Construction We create comprehensive peptide or antibody libraries for traditional phage display techniques.
Peptide Synthesis & Modification Custom synthesis and modification of peptides for further research and development.
Antibody Development Platforms Full-spectrum antibody discovery, optimization, and production services.

Creative Biolabs' mirror-image phage display service provides a unique and powerful tool for discovering stable, high-affinity peptides and antibodies for therapeutic and diagnostic applications. With our extensive experience, cutting-edge technologies, and personalized approach, we can help accelerate your research and development efforts. Whether you're looking for D-peptide drug candidates, high-affinity binders, or optimized antibodies, our MIPD service is the solution you need. Contact us today to start your next project.

Published Data2,3

The transformative potential of mirror-image phage display is powerfully illustrated in a landmark study focused on cancer immunotherapy, where researchers successfully developed a potent D-peptide antagonist against the critical immune checkpoint interaction between TIGIT and its ligand, PVR. The rationale for the study was clear: while the TIGIT/PVR pathway is a key suppressor of T-cell anti-tumor activity and a prime therapeutic target, conventional antibody-based inhibitors face significant hurdles, including high manufacturing costs and poor penetration into the dense tumor microenvironment. To overcome these challenges, the research team employed our mirror-image display strategy. They began with the total chemical synthesis of the D-enantiomer of the TIGIT protein, creating a perfect mirror image of the natural target. This synthetic D-TIGIT was then used as a molecular bait to screen a commercial L-peptide phage display library, isolating several high-affinity binders. The most promising L-peptide sequence was then used as a blueprint to synthesize its D-peptide counterpart, named DPPA-1 chemically. The subsequent validation results underscored the success of this approach. In vitro assays confirmed that DPPA-1 specifically and effectively blocked the binding of TIGIT to PVR, which robustly enhanced T-cell activation and the secretion of anti-tumor cytokines. More impressively, when evaluated in a murine melanoma model, DPPA-1 demonstrated significant therapeutic efficacy, markedly inhibiting tumor growth and extending the survival of the subjects. This study brilliantly highlights the core advantages of the technology. As a D-peptide, DPPA-1 is inherently resistant to the enzymatic degradation that rapidly clears natural L-peptides, ensuring superior stability in vivo. Furthermore, its small molecular size enhances tissue and tumor permeability compared to bulky antibodies, addressing a major delivery obstacle. This combination of high stability and deep tissue penetration opens new possibilities for creating more effective, next-generation immunotherapies, including potential oral formulations or long-acting injectables.

FAQs

Q: What is a mirror-image in biology?

A: In biology, a mirror-image refers to the opposite enantiomer of a molecule, such as D-peptides, which are the mirror image of L-peptides. These mirror-image molecules maintain the same molecular structure but have different properties, such as greater resistance to enzymatic breakdown.

Q: What is mirror-image phage display?

A: Mirror-image phage display is a technology that screens L-peptides against a synthesized D-form of your biological target. The identified L-peptides are then chemically synthesized as D-peptides, which are more stable, protease-resistant, and ideal for therapeutic and diagnostic development.

Q: What types of molecules can be used as targets?

A: MIPD is most effective for peptide and small protein targets that can be chemically synthesized in D-form. Common targets include epitopes of viral proteins, receptor-binding domains, or disease-related short peptides. Larger proteins are not suitable due to synthesis limitations.

Q: Is mirror-image phage display suitable for vaccine or immunogen discovery?

A: MIPD is not typically used for vaccines since D-peptides are non-immunogenic. However, it's excellent for creating stable, non-immunogenic ligands that mimic antigenic surfaces and may act as decoys or blockers in infectious disease or autoimmunity research.

References:

  1. Sadraeian, Mohammad, et al. "Phage display technology in biomarker identification with emphasis on non-cancerous diseases." Molecules 29.13 (2024): 3002. https://doi.org/10.3390/molecules29133002
  2. Sui, Xinghua, et al. "Peptide drugs: a new direction in cancer immunotherapy." Cancer Biology & Medicine 21.3 (2023): 198. https://doi.org/10.20892/j.issn.2095-3941.2023.0297
  3. Distributed under Open Access license CC BY 4.0, without modification.

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