scholarly journals A Novel Cu(II)-Binding Peptide Identified by Phage Display Inhibits Cu2+-Mediated Aβ Aggregation

2021 ◽  
Vol 22 (13) ◽  
pp. 6842
Author(s):  
Xiaoyu Zhang ◽  
Xiancheng Zhang ◽  
Manli Zhong ◽  
Pu Zhao ◽  
Chuang Guo ◽  
...  
Keyword(s):  
Phage Display ◽  
Metal Ion ◽  
Amyloid Β ◽  
Amyloid Β Peptide ◽  
Ion Distribution ◽  
Phage Display Technology ◽  
Binding Peptide ◽  
Aβ Aggregation ◽  
Display Technology

Copper (Cu) has been implicated in the progression of Alzheimer’s disease (AD), and aggregation of Cu and amyloid β peptide (Aβ) are considered key pathological features of AD. Metal chelators are considered to be potential therapeutic agents for AD because of their capacity to reduce metal ion-induced Aβ aggregation through the regulation of metal ion distribution. Here, we used phage display technology to screen, synthesize, and evaluate a novel Cu(II)-binding peptide that specifically blocked Cu-triggered Aβ aggregation. The Cu(II)-binding peptide (S-A-Q-I-A-P-H, PCu) identified from the phage display heptapeptide library was used to explore the mechanism of PCu inhibition of Cu2+-mediated Aβ aggregation and Aβ production. In vitro experiments revealed that PCu directly inhibited Cu2+-mediated Aβ aggregation and regulated copper levels to reduce biological toxicity. Furthermore, PCu reduced the production of Aβ by inhibiting Cu2+-induced BACE1 expression and improving Cu(II)-mediated cell oxidative damage. Cell culture experiments further demonstrated that PCu had relatively low toxicity. This Cu(II)-binding peptide that we have identified using phage display technology provides a potential therapeutic approach to prevent or treat AD.

scholarly journals Novel Blood–Brain Barrier Shuttle Peptides Discovered through the Phage Display Method

Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 874 ◽  
Author(s):  
Petra Majerova ◽  
Jozef Hanes ◽  
Dominika Olesova ◽  
Jakub Sinsky ◽  
Emil Pilipcinec ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Phage Display ◽  
Blood Brain Barrier ◽  
Brain Barrier ◽  
Phage Display Technology ◽  
Blood Brain ◽  
Display Technology ◽  
The Brain

Delivery of therapeutic agents into the brain is a major challenge in central nervous system drug development. The blood–brain barrier (BBB) prevents access of biotherapeutics to their targets in the central nervous system and, therefore, prohibits the effective treatment of many neurological disorders. To find blood–brain barrier shuttle peptides that could target therapeutics to the brain, we applied a phage display technology on a primary endothelial rat cellular model. Two identified peptides from a 12 mer phage library, GLHTSATNLYLH and VAARTGEIYVPW, were selected and their permeability was validated using the in vitro BBB model. The permeability of peptides through the BBB was measured by ultra-performance liquid chromatography-tandem mass spectrometry coupled to a triple-quadrupole mass spectrometer (UHPLC-MS/MS). We showed higher permeability for both peptides compared to N–C reversed-sequence peptides through in vitro BBB: for peptide GLHTSATNLYLH 3.3 × 10−7 cm/s and for peptide VAARTGEIYVPW 1.5 × 10−6 cm/s. The results indicate that the peptides identified by the in vitro phage display technology could serve as transporters for the administration of biopharmaceuticals into the brain. Our results also demonstrated the importance of proper BBB model for the discovery of shuttle peptides through phage display libraries.

In vitro effect of a thrombin inhibition peptide selected by phage display technology

2002 ◽  
Vol 107 (6) ◽  
pp. 365-371 ◽  
Author(s):  
Muriel S Meiring ◽  
Derek Litthauer ◽  
Jolan Hársfalvi ◽  
Veronica van Wyk ◽  
Philip N Badenhorst ◽  
...  
Keyword(s):  
Phage Display ◽  
Phage Display Technology ◽  
Thrombin Inhibition ◽  
Display Technology ◽  
Vitro Effect

scholarly journals A novel, stable, helical scaffold as an alternative binder - construction of phage display libraries.

2012 ◽  
Vol 59 (3) ◽  
Author(s):  
Anna Cyranka-Czaja ◽  
Jacek Otlewski
Keyword(s):  
Phage Display ◽  
Measles Virus ◽  
In Vitro Selection ◽  
Biophysical Properties ◽  
Phage Display Technology ◽  
High Affinity ◽  
Promising Tool ◽  
Phage Display Libraries ◽  
Display Technology

Specific, high affinity binding macromolecules are of great importance for biomedical and biotechnological applications. The most popular classical antibody-based molecules have recently been challenged by alternative scaffolds with desirable biophysical properties. Phage display technology applied to such scaffolds allows generation of potent affinity reagents by in vitro selection. Here, we report identification and characterization of a novel helical polypeptide with advantageous biophysical properties as a template for construction of phage display libraries. A three-helix bundle structure, based on Measles virus phosphoprotein P shows a very favourable stability and solubility profile. We designed, constructed and characterized six different types of phage display libraries based on the proposed template. Their functional size of over 10(9) independent clones, balanced codon bias and decent display level are key parameters attesting to the quality and utility of the libraries. The new libraries are a promising tool for isolation of high affinity binders based on a small helical scaffold which could become a convenient alternative to antibodies.

scholarly journals Identification of Novel Single-Domain Antibodies against FGF7 Using Phage Display Technology

2017 ◽  
Vol 23 (2) ◽  
pp. 193-201
Author(s):  
Behzad Jafari ◽  
Maryam Hamzeh-Mivehroud ◽  
Ali A. Moosavi-Movahedi ◽  
Siavoush Dastmalchi
Keyword(s):  
Growth Factor ◽  
Phage Display ◽  
Fibroblast Growth Factor ◽  
Single Domain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fibroblast Growth ◽  
Phage Display Technology ◽  
Display Technology ◽  
Single Domain Antibodies

Fibroblast growth factor 7 (FGF7) is a member of the fibroblast growth factor (FGF) family of proteins. FGF7 is of stromal origin and produces a paracrine effect on epithelial cells. In the current investigation, we aimed to identify new single-domain antibodies (sdAbs) against FGF7 using phage display technology. The vector harboring the codon-optimized DNA sequence for FGF7 protein was transformed into Escherichia coli BL21 (DE3) pLysS, and then the protein was expressed at the optimized condition. Enzyme-linked immunosorbent assay, circular dichroism spectropolarimetry, and in vitro scratch assay experiments were used to confirm the proper folding and functionality of the purified FGF7 protein. The purity of the produced FGF7 was 92%, with production yield of 3.5 mg/L of culture. Panning against the purified FGF7 was performed, and the identified single-domain antibodies showed significant affinity. Further investigation on one of the selected sdAb displaying phage clones showed concentration-dependent binding to FGF7. The selected sdAb can be used for developing novel tumor-suppressing agents where inhibition of FGF7 is required.

A Cationic Gallium Phthalocyanine Inhibits Amyloid β Peptide Fibril Formation

2020 ◽  
Vol 17 (7) ◽  
pp. 589-600
Author(s):  
Shatera Tabassum ◽  
Abdullah Md. Sheikh ◽  
Shozo Yano ◽  
Takahisa Ikeue ◽  
Shingo Mitaki ◽  
...  
Keyword(s):  
Near Infrared ◽  
Amyloid Β ◽  
Neuronal Cell ◽  
Fibril Formation ◽  
Lag Phase ◽  
Slice Culture ◽  
Amyloid Β Peptide ◽  
Protective Effects ◽  
Aβ Aggregation

Background: Amyloid β (Aβ) peptide deposition is considered as the main cause of Alzheimer’s disease (AD). Previously, we have shown that a Zn containing neutral phthalocyanine (Zn-Pc) inhibits Aβ fibril formation. Objective: The objective of this study is to investigate the effects of a cationic gallium containing Pc (GaCl-Pc) on Aβ fibril formation process. Methods and Results: Aβ fibril formation was induced by incubating synthetic Aβ peptides in a fibril forming buffer, and the amount of fibril was evaluated by ThT fluorescence assay. GaCl-Pc dosedependently inhibited both Aβ1-40 and Aβ1-42 fibril formation. It mainly inhibited the elongation phase of Aβ1-42 fibril formation kinetics, but not the lag phase. Western blotting results showed that it did not inhibit its oligomerization process, rather increased it. Additionally, GaCl-Pc destabilized preformed Aβ1- 42 fibrils dose-dependently in vitro condition, and decreased Aβ levels in the brain slice culture of APP transgenic AD model mice (J20 strain). Near-infrared scanning results showed that GaCl-Pc had the ability to bind to Aβ1-42. MTT assay demonstrated that GaCl-Pc did not have toxicity towards a neuronal cell line (A1) in culture rather, showed protective effects on Aβ-induced toxicity. Moreover, it dosedependently decreased Aβ-induced reactive oxygen species levels in A1 culture. Conclusion: Thus, our result demonstrated that GaCl-Pc decreased Aβ aggregation and destabilized the preformed fibrils. Since cationic molecules show a better ability to cross the blood-brain barrier, cationic GaCl-Pc could be important for the therapy of AD.

scholarly journals Challenges in Optimizing a Prostate Carcinoma Binding Peptide, Identified through the Phage Display Technology

Molecules ◽  
2011 ◽  
Vol 16 (2) ◽  
pp. 1559-1578 ◽  
Author(s):  
Vasileios Askoxylakis ◽  
Sabine Zitzmann-Kolbe ◽  
Frederic Zoller ◽  
Annette Altmann ◽  
Annette Markert ◽  
...  
Keyword(s):  
Phage Display ◽  
Prostate Carcinoma ◽  
Phage Display Technology ◽  
Binding Peptide ◽  
Display Technology

scholarly journals Metal ion coordination delays amyloid-β peptide self-assembly by forming an aggregation–inert complex

2020 ◽  
Vol 295 (21) ◽  
pp. 7224-7234 ◽  
Author(s):  
Cecilia Wallin ◽  
Jüri Jarvet ◽  
Henrik Biverstål ◽  
Sebastian Wärmländer ◽  
Jens Danielsson ◽  
...  
Keyword(s):  
Metal Ions ◽  
Self Assembly ◽  
Metal Ion ◽  
Amyloid Β ◽  
Bound State ◽  
Ion Binding ◽  
Amyloid Β Peptide ◽  
Aβ Peptide ◽  
Metal Ion Binding

A detailed understanding of the molecular pathways for amyloid-β (Aβ) peptide aggregation from monomers into amyloid fibrils, a hallmark of Alzheimer's disease, is crucial for the development of diagnostic and therapeutic strategies. We investigate the molecular details of peptide fibrillization in vitro by perturbing this process through addition of differently charged metal ions. Here, we used a monovalent probe, the silver ion, that, similarly to divalent metal ions, binds to monomeric Aβ peptide and efficiently modulates Aβ fibrillization. On the basis of our findings, combined with our previous results on divalent zinc ions, we propose a model that links the microscopic metal-ion binding to Aβ monomers to its macroscopic impact on the peptide self-assembly observed in bulk experiments. We found that substoichiometric concentrations of the investigated metal ions bind specifically to the N-terminal region of Aβ, forming a dynamic, partially compact complex. The metal-ion bound state appears to be incapable of aggregation, effectively reducing the available monomeric Aβ pool for incorporation into fibrils. This is especially reflected in a decreased fibril-end elongation rate. However, because the bound state is significantly less stable than the amyloid state, Aβ peptides are only transiently redirected from fibril formation, and eventually almost all Aβ monomers are integrated into fibrils. Taken together, these findings unravel the mechanistic consequences of delaying Aβ aggregation via weak metal-ion binding, quantitatively linking the contributions of specific interactions of metal ions with monomeric Aβ to their effects on bulk aggregation.

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