Phage Display and Synthetic Peptides as Promising Biotechnological Tools for the Serological Diagnosis of Leprosy
نویسندگان
چکیده
BACKGROUND The diagnosis of leprosy is primarily based on clinical manifestations, and there is no widely available laboratory test for the early detection of this disease, which is caused by Mycobacterium leprae. In fact, early detection and treatment are the key elements to the successful control of leprosy. METHODOLOGY/PRINCIPAL FINDINGS Peptide ligands for antibodies from leprosy patients were selected from phage-displayed peptide libraries. Three peptide sequences expressed by reactive phage clones were chemically synthesized. Serological assays that used synthetic peptides were evaluated using serum samples from leprosy patients, household contacts (HC) of leprosy patients, tuberculosis patients and endemic controls (EC). A pool of three peptides identified 73.9% (17/23) of multibacillary (MB) leprosy patients using an enzyme-linked immunosorbent assay (ELISA). These peptides also showed some seroreactivities to the HC and EC individuals. The peptides were not reactive to rabbit polyclonal antisera against the different environmental mycobacteria. The same peptides that were conjugated to the carrier protein bovine serum albumin (BSA) induced the production of antibodies in the mice. The anti-peptide antibodies that were used in the Western blotting analysis of M. leprae crude extracts revealed a single band of approximately 30 kDa in one-dimensional electrophoresis and four 30 kDa isoforms in the two-dimensional gel. The Western blotting data indicated that the three peptides are derived from the same bacterial protein. CONCLUSIONS/SIGNIFICANCE These new antigens may be useful in the diagnosis of MB leprosy patients. Their potentials as diagnostic reagents must be more extensively evaluated in future studies using a large panel of positive and negative sera. Furthermore, other test approaches using peptides should be assessed to increase their sensitivity and specificity in detecting leprosy patients. We have revealed evidence in support of phage-displayed peptides as promising biotechnological tools for the design of leprosy diagnostic serological assays.
منابع مشابه
Identification 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 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 کاملDiscovery of Novel Peptidomimetics for Brain-Derived Neurotrophic Factor using Phage Display Technology
Brain-Derived Neurotrophic Factor (BDNF) is a neuroprotectant candidate for neurodegenerative diseases. However, there are several clinical concerns about its therapeutic applications. In the current study, we selected BDNF-mimicking small peptides from phage-displayed peptide library as alternative molecules to the clinical challenges. The peptide library was screened against BDNF receptor (Ne...
متن کاملPuzzling Peptides from a Phage Display Library
The commercial availability of random peptide libraries displayed on the M13 phage is increasing their use forstudies on epitope identification, enzyme inhibitors, receptor ligands, etc. In this study two experimentswhere planned for selection of peptides. First with sheep antibodies, the positive selector was IgG, preparedon Protein G column from a pool of 11 sheeps immunized...
متن کاملExpression and Purification of Brucella-Specific Nanobodies
Background: Brucellosis is still considered as one of the major zoonosis afflicting Syrian health and economy. This disease is caused by members of the genus Brucella which are gram-negative bacteria living facultatively within mammalian cells during infection. Objectives: In this paper, a strategy was developed to introduce a new generation of binders called Nanobodies (Nbs) in our combat aga...
متن کامل