Infection and Drug Resistance (Aug 2022)
Bioprospecting by Phage Display of Mimetic Peptides of Chlamydia trachomatis for Use in Laboratory Diagnosis
Abstract
Larissa Silva de Freitas,1,* Maria Alice Freitas Queiroz,1,* Luiz Fernando Almeida Machado,1 Antonio Carlos Rosário Vallinoto,1 Marluísa de Oliveira Guimarães Ishak,1 Fabiana de Almeida Araújo Santos,2 Luiz Ricardo Goulart2 ,† Ricardo Ishak1 1Laboratory of Virology, Biological Sciences Institute, Federal University of Pará, Belém, Pará, Brazil; 2Laboratory of Nanobiotechnology, Genetics and Biochemistry Institute, Federal University of Uberlândia, Uberlândia, Minas Gerais, Brazil†Luiz Ricardo Goulart passed away on 24/10/2021*These authors contributed equally to this workCorrespondence: Maria Alice Freitas Queiroz, Laboratory of Virology, Biological Sciences Institute, Federal University of Pará, Belém, Pará, Brazil, Tel +55 91 3201-7587, Email [email protected]: Chlamydia trachomatis infection is a major public health problem and the most common sexually transmitted infection in the world. Although highly prevalent, 70% to 80% of cases are asymptomatic and undiagnosed.Purpose: To overcome some limitations in terms of rapid diagnosis, phage display technology was used to bioprospect peptide mimetics of C. trachomatis immunoreactive and immunogenic antigens to be selected for the production of synthetic peptides.Methods: Initially, IgG from 22 individuals with C. trachomatis and 30 negative controls was coupled to G protein magnetic beads. The phage display technique consisted of biopanning, genetic sequencing, bioinformatics analysis and phage ELISA.Results: Clones G1, H5, C6 and H7 were selected for testing with individual samples positive and negative for C. trachomatis. Reactions were statistically significant (p 0.8. One-dimensional analysis with C. trachomatis components indicated that the G1 clone aligned with cell wall-associated hydrolase domain-containing protein, the H5 clone aligned with glycerol-3-phosphate acyltransferase PlsX protein, the C6 clone aligned with a transposase and inactivated derivatives, and the H7 clone aligned with GTP-binding protein. Molecular modeling and three-dimensional analysis indicated the best fit of the four clones with a protein known as chlamydial protease/proteasome-like activity factor (CPAF), an important virulence factor of the bacterium.Conclusion: The peptides produced by phage display are related to the metabolic pathways of C. trachomatis, indicating that they can be used to understand the pathogenesis of the infection. Because of their high sensitivity and AUC values, the peptides present considerable potential for use in platforms for screening C. trachomatis infections.Keywords: C. trachomatis, phage display, mimetic peptides, CPAF, screening tests
