A Novel Screening System for Claudin Binder Using Baculoviral Display
نویسندگان
چکیده
Recent progress in cell biology has provided new insight into the claudin (CL) family of integral membrane proteins, which contains more than 20 members, as a target for pharmaceutical therapy. Few ligands for CL have been identified because it is difficult to prepare CL in an intact form. In the present study, we developed a method to screen for CL binders by using the budded baculovirus (BV) display system. CL4-displaying BV interacted with a CL4 binder, the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE), but it did not interact with C-CPE that was mutated in its CL4-binding region. C-CPE did not interact with BV and CL1-displaying BV. We used CL4-displaying BV to select CL4-binding phage in a mixture of a scFv-phage and C-CPE-phage. The percentage of C-CPE-phage in the phage mixture increased from 16.7% before selection to 92% after selection, indicating that CL-displaying BV may be useful for the selection of CL binders. We prepared a C-CPE phage library by mutating the functional amino acids. We screened the library for CL4 binders by affinity to CL4-displaying BV, and we found that the novel CL4 binders modulated the tight-junction barrier. These findings indicate that the CL-displaying BV system may be a promising method to produce a novel CL binder and modulator.
منابع مشابه
One-Step Recovery of scFv Clones from High-Throughput Sequencing-Based Screening of Phage Display Libraries Challenged to Cells Expressing Native Claudin-1
Expanding the availability of monoclonal antibodies interfering with hepatitis C virus infection of hepatocytes is an active field of investigation within medical biotechnologies, to prevent graft reinfection in patients subjected to liver transplantation and to overcome resistances elicited by novel antiviral drugs. In this paper, we describe a complete pipeline for screening of phage display ...
متن کاملTitle: Creation of a claudin-2 binder and its tight-junction-modulating activity in a human intestinal model
متن کامل
Enhanced Bioadsorption of Cadmium and Nickel by E. coli Displaying A Metal Binding Motif Using CS3 Fimbriae
Display of peptides on the surface of bacteria offers many new and exciting applications in biotechnology. Fimbriae is a good candidate for epitope display on the surface of bacteria. The potential of CS3 fimbriae of enterotoxigenic E. coli as a display system has been investigated. A novel cell surface display system with metal binding property was developed by using CS3 fimbriae. Short metal ...
متن کاملIdentification of a Novel Tumor-Binding Peptide for Lung Cancer Through in-vitro Panning
Tumor-targeted therapies are playing growing roles in cancer research. The exploitation of these powerful therapeutic modalities largely depends on the discovery of tumor-targeting ligands. Phage display has proven a promising high throughput screening tool for the identification of novel specific peptides with high binding affinity to cancer cells. In the present study, we describe the use of ...
متن کاملImproved Procedure for Screening Expression Libraries for Novel Autoantigens
The standard method for immunoscreening of a cDNA expression library is time-consuming becauseof the production of a large proportion of false positives during the first and second round of screening.This problem is more important when a sensitive chemiluminescence detection system is used. Due tothe high sensitivity of the detection system, there is a need to avoid false posi...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 6 شماره
صفحات -
تاریخ انتشار 2011