Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis
Ana Alice Maia Gonçalves, Anna Júlia Ribeiro, Carlos Ananias Aparecido Resende, Carolina Alves Petit Couto, Isadora Braga Gandra, Isabelle Caroline Dos Santos Barcelos, Jonatas Oliveira da Silva, Juliana Martins Machado, Kamila Alves Silva, Líria Souza Silva, Michelli dos Santos, Lucas da Silva Lopes, Mariana Teixeira de Faria, Sabrina Paula Pereira, Sandra Rodrigues Xavier, Matheus Aragão, Mayron Antonio Candida-Puma, Izadora Oliveira, Amanda Araújo Souza, Laís Moreira Nogueira, Mariana Campos-da-Paz, Eduardo Antônio Ferraz Coelho, Rodolfo Cordeiro Giunchetti, Sônia Maria de Freitas, Miguel Á. Chávez‐Fumagalli, Ronaldo Alves Pinto Nagem, Alexsandro Sobreira Galdino
Abstract
Recombinant multiepitope proteins (RMPs) are a promising alternative for application in diagnostic tests and, given their wide application in the most diverse diseases, this review article aims to survey the use of these antigens for diagnosis, as well as discuss the main points surrounding these antigens. RMPs usually consisting of linear, immunodominant, and phylogenetically conserved epitopes, has been applied in the experimental diagnosis of various human and animal diseases, such as leishmaniasis, brucellosis, cysticercosis, Chagas disease, hepatitis, leptospirosis, leprosy, filariasis, schistosomiasis, dengue, and COVID-19. The synthetic genes for these epitopes are joined to code a single RMP, either with spacers or fused, with different biochemical properties. The epitopes' high density within the RMPs contributes to a high degree of sensitivity and specificity. The RMPs can also sidestep the need for multiple peptide synthesis or multiple recombinant proteins, reducing costs and enhancing the standardization conditions for immunoassays. Methods such as bioinformatics and circular dichroism have been widely applied in the development of new RMPs, helping to guide their construction and better understand their structure. Several RMPs have been expressed, mainly using the Escherichia coli expression system, highlighting the importance of these cells in the biotechnological field. In fact, technological advances in this area, offering a wide range of different strains to be used, make these cells the most widely used expression platform. RMPs have been experimentally used to diagnose a broad range of illnesses in the laboratory, suggesting they could also be useful for accurate diagnoses commercially. On this point, the RMP method offers a tempting substitute for the production of promising antigens used to assemble commercial diagnostic kits.