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Multivalent pIX Display based M13 Phage Library Construction Service

Background Service FAQ Resources

Phage display screening plays a key role in the identification of a multitude of antibody specificities, but identifying optimal candidates still remains a challenge. Creative Biolabs provides high-quality service in phage display antibody library construction and screening. Our scientists developed a pIX combinatorial display system, combining with the use of an engineered helper phage. This approach provides a generic alternative route for obtaining good binders with higher thermostability and affinity. Creative Biolabs is pleased to introduce our pIX library construction and screening service to our global clients.

Background

Phage display has largely replaced hybridoma technology for therapeutic monoclonal antibody development. As there is a growing need for improved targeted therapeutic intervention, phage display technology has been extensively modified and improved since its introduction three decades ago. By far, pIII is the dominating display capsid used in antibody display and selection. However, the use of pIII is associated with well-documented negative effects on selection performance, such as clone-dependent reduced phage infectivity and propagation, which cause repertoire bias. Thus, the limitations in pIII display technology motivate the search for further improvements. Several reports have explored pIX as an alternative antibody display scaffold. pIX and pIII are located at opposite tips on the virion, and the two proteins differ in their mechanisms of translocation to the E. coli inner membrane prior to phage assembly. pIX lacks a signal sequence. It does not undergo post-translational processing and appears to depend on YidC for periplasmic targeting. Therefore, signal sequences are added to facilitate antibody display, but later reports are conflicting with respect to how well such pIX display performs in comparison to display on pIII.

Overview of pIX display routes (Høydahl et al. 2016).Fig.1 Overview of pIX display routes.1

Multivalent pIX Display based M13 Phage Library Construction Service

Creative Biolabs has developed a multivalent pIX display system with the high hit-rate of specific clones in diverse antibody library selection, avoiding the use of a signal sequence to the fusion protein. This approach uses an engineered helper phage known to improve antibody pIX display and retrieval. Multivalent pIX phage display screening is suitable for distinct and improved antibody properties.

Key advantages including but not limited to:


Creative Biolabs' scientists have extensive experience in pIX phage display library construction and screening. Our scientists have demonstrated that this approach translates into substantially improved retrieval of desired specificities with favorable biophysical properties in de novo selection. We are confident in providing clients with the best service at the most competitive cost.

For more detailed information, please feel free to contact us or directly sent us an inquiry.

Other optional M13 phage library construction service:

Reference
  1. Hoydahl, L. S., et al. "Multivalent pIX phage display selects for distinct and improved antibody properties. Sci Rep. 2016; 6: 39066." Distributed under Open Access license CC BY 4.0, without modification.

FAQ

  1. What is a Multivalent pIX Display-based M13 Phage Library, and how does it differ from other phage display systems?

    The Multivalent pIX Display-based M13 Phage Library is a specialized variant of the M13 phage display system where peptides or proteins are displayed on the pIX protein of the M13 phage, which is located at the phage's end opposite to the pIII or pVIII proteins. This multivalent display allows for the presentation of multiple copies of a peptide on the same phage particle, potentially enhancing the binding affinity and avidity for the target. This system is particularly useful for selecting peptides that require multivalent interactions for effective binding.

  2. What are the primary applications of the Multivalent pIX Display-based M13 Phage Library in research?

    The Multivalent pIX Display-based M13 Phage Library is used in drug discovery, vaccine development, and the study of protein-protein interactions. It is particularly valuable for identifying ligands that require multivalent binding to achieve high affinity and specificity. This system is also used to explore and develop peptide-based therapeutics, where multiple weak interactions can collectively result in strong target binding.

  3. How does the multivalent nature of the pIX display enhance the selection process in phage display libraries?

    The multivalent nature of the pIX display means that multiple copies of a peptide are presented on the surface of the phage, which can significantly enhance binding through cooperative interactions. This increased binding strength, known as avidity, is particularly beneficial when selecting peptides that need to engage with multiple binding sites on a target, leading to more robust and specific selections compared to monovalent displays.

  4. What are the key differences between pIX and pIII/pVIII display platforms in M13 phage libraries?

    In M13 phage display, the pIII and pVIII proteins are commonly used for displaying peptides. The pIX display differs in that it is located at the opposite end of the phage from pIII and is capable of presenting peptides in a multivalent format. While pIII typically displays up to five copies of a peptide and pVIII displays thousands, pIX allows for a moderate number of peptide copies (typically 5-10 per phage), balancing the benefits of multivalency without overcrowding the displayed peptides, which can occur with pVIII.

  5. How is the Multivalent pIX Display-based M13 Phage Library constructed, and what are its advantages?

    The Multivalent pIX Display-based M13 Phage Library is constructed by inserting DNA sequences encoding the desired peptides into the gene encoding the pIX protein of the M13 phage. This insertion allows the peptides to be displayed on the pIX protein at the end of the phage particle. The main advantage of this system is its ability to display peptides multivalently, increasing the likelihood of strong interactions with the target and improving the overall selection process, especially for targets that require multivalent engagement.

  6. How is the Multivalent pIX Display-based M13 Phage Library utilized in drug discovery?

    In drug discovery, the Multivalent pIX Display-based M13 Phage Library is used to identify peptide ligands that can bind to therapeutic targets with high specificity and affinity. The multivalent display of peptides on the pIX protein allows for the selection of ligands that may interact with multiple binding sites on a target, enhancing the therapeutic potential of the selected peptides. This approach is particularly useful for identifying inhibitors or activators of complex protein-protein interactions.

  7. What factors influence the success of selections using the Multivalent pIX Display-based M13 Phage Library?

    The quality and diversity of the library, the nature of the target, and the selection conditions could influence the success of selections using the Multivalent pIX Display-based M13 Phage Library. The density of the displayed peptides must be optimized to avoid steric hindrance while maintaining effective multivalent interactions. Additionally, the washing and elution steps during biopanning must be carefully controlled to enrich for the highest affinity binders without losing potentially valuable candidates.

Resources

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