Immunodiagnosis of Canine Visceral Leishmaniasis Using Mimotope Peptides Selected from Phage Displayed Combinatorial Libraries

ELISA and RIFI are currently used for serodiagnosis of canine visceral leishmaniasis (CVL). The accuracy of these tests is controversial in endemic areas where canine infections byTrypanosoma cruzimay occur. We evaluated the usefulness of synthetic peptides that were selected through phage display technique in the serodiagnosis of CVL. Peptides were chosen based on their ability to bind to IgGs purified from infected dogs pooled sera. We selected three phage clones that reacted only with those IgGs. Peptides were synthesized, polymerized with glutaraldehyde, and used as antigens in ELISA assays. Each individual peptide or a mix of them was reactive with infected dogs serum. The assay was highly sensitive and specific when compared to solubleLeishmaniaantigen that showed cross-reactivity with anti-T. cruziIgGs. Our results demonstrate that phage display technique is useful for selection of peptides that may represent valuable synthetic antigens for an improved serodiagnosis of CVL.

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The phage display technique and transfusion medicine

Transfusion ◽

10.1046/j.1537-2995.2001.41010001.x ◽

2001 ◽

Vol 41 (1) ◽

pp. 1-3 ◽

Cited By ~ 3

Author(s):

Petr Jarolim

Keyword(s):

Phage Display ◽

Transfusion Medicine ◽

Phage Display Technique ◽

Display Technique

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Performance of recombinant proteins in diagnosis and differentiation of canine visceral leishmaniasis infected and vaccinated dogs

European Journal of Microbiology and Immunology ◽

10.1556/1886.2020.00018 ◽

2020 ◽

Vol 10 (3) ◽

pp. 165-171

Author(s):

Ingrid E. Pereira ◽

Kyssia P. Silva ◽

Laura M. Menegati ◽

Aimara C. Pinheiro ◽

Elaine A. O. Assunção ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Recombinant Proteins ◽

Serological Test ◽

High Sensitivity ◽

High Specificity ◽

Cross Reactivity ◽

Reservoir Host ◽

Superior Performance ◽

Canine Visceral Leishmaniasis ◽

Roc Curve Analysis

AbstractControl of canine visceral leishmaniasis (CVL), a major zoonotic disease in Brazil and many other tropical and subtropical countries, remains difficult as an accurate and reliable diagnosis is still missing. In endemic regions, infected dogs are the main parasitic reservoir host of human Visceral leishmaniasis (VL) infection. Vaccination of dogs against Leishmania infection constitutes an important strategy to prevent or to better control CVL, thus, a serological test that can discriminate between antibodies induced by immunization versus infection is highly desirable in order to improve and simplify diagnosis. Here, four recombinant proteins were evaluated for their ability to detect and differentiate between dogs that are infected with Leishmania or have been immunized with the anti-Leishmania vaccine Leish-Tec®. Receiver operating characteristic (ROC) curve analysis of the four Leishmania-specific IgG ELISA revealed superior performance of rK28, followed by rKLO8, rK39 and rLb6H. The rK28-based ELISA revealed not only the best accuracy against CVL, but also the lowest cross-reactivity with sera from Leish-Tec® immunized dogs. Our data show that the rK28-based ELISA is highly suitable for CVL screening as it shows high sensitivity with simultaneous low cross-reactivity. Further, the high specificity of the rKLO8 indicates its suitability for the confirmation of CVL diagnosis.

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Development of Human Antibody Inhibiting RNase H Activity of Polymerase of Hepatitis B Virus Using Phage Display Technique

Immune Network ◽

10.4110/in.2004.4.1.16 ◽

2004 ◽

Vol 4 (1) ◽

pp. 16

Author(s):

Seong-Rak Lee ◽

Eun-Kyoung Song ◽

Young-Joo Jeong ◽

Young-Yi Lee ◽

Ik-Jung Kim ◽

...

Keyword(s):

Hepatitis B Virus ◽

Hepatitis B ◽

Phage Display ◽

Rnase H ◽

Human Antibody ◽

Phage Display Technique ◽

B Virus ◽

Display Technique

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A Novel Method forin SituDetection of Hydrolyzable Casein Fragments in a Cheese Matrix by Antibody Phage Display Technique and CLSM

Journal of Agricultural and Food Chemistry ◽

10.1021/jf901026m ◽

2009 ◽

Vol 57 (21) ◽

pp. 10049-10054 ◽

Cited By ~ 3

Author(s):

Zhi Duan ◽

Dagmar Adeline Brüggemann ◽

Henrik Siegumfeldt

Keyword(s):

Phage Display ◽

Phage Display Technique ◽

Antibody Phage ◽

Antibody Phage Display ◽

Novel Method ◽

Display Technique

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Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications

Clinical and Vaccine Immunology ◽

10.1128/cvi.00583-13 ◽

2013 ◽

Vol 21 (1) ◽

pp. 96-106 ◽

Cited By ~ 18

Author(s):

Lourena E. Costa ◽

Mayara I. S. Lima ◽

Miguel A. Chávez-Fumagalli ◽

Daniel Menezes-Souza ◽

Vivian T. Martins ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Phage Display ◽

Leishmania Infantum ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Phage Display Technology ◽

Predictive Values ◽

Content Type ◽

Phage Elisa

ABSTRACTVisceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy andTrypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected withLeishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n= 31) compared to those from vaccinated dogs (n= 21), experimentally infected dogs with cross-reactive parasites (n= 23), and healthy controls (n= 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no cross-reactivity withT. cruzi- orEhrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes ofL. infantumantigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs.

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Phage-based Nanomedicines as New Immune Therapeutic Agents for Breast Cancer

Current Pharmaceutical Design ◽

10.2174/1381612824666180327152117 ◽

2018 ◽

Vol 24 (11) ◽

pp. 1195-1203 ◽

Cited By ~ 3

Author(s):

Atefeh Arab ◽

Rezvan Yazdian Robati ◽

Jessica Nicastro ◽

Roderick Slavcev ◽

Javad Behravan

Keyword(s):

Breast Cancer ◽

Phage Display ◽

Biomedical Applications ◽

Therapeutic Agents ◽

Molecular Method ◽

Specific Antibodies ◽

Phage Display Technique ◽

Specific Antigens ◽

Protein Libraries ◽

Display Technique

Despite years of investigation, breast cancer remains a major cause of death worldwide. Phage display is a powerful molecular method in which peptide and protein libraries can be displayed via genetic fusions on the surface of phages. This approach has tremendous potential for biomedical applications and has already facilitated the discovery of specific antibodies, specific antigens, and peptides with potential roles in the diagnosis and treatment of malignancies including breast cancer. In this review, we discuss the new and the latest advancements in the applications of the phage display technique in the provision of immune therapeutics for breast cancer.

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