Rapid Identification of Emerging Pathogens: Coronavirus

New surveillance approach can analyze >900 polymerase chain reactions per day

An update on novel approaches for diagnosis and treatment of SARS-CoV-2 infection

The ongoing pandemic of coronavirus disease 2019 (COVID-19) has made a serious public health and economic crisis worldwide which united global efforts to develop rapid, precise, and cost-efficient diagnostics, vaccines, and therapeutics. Numerous multi-disciplinary studies and techniques have been designed to investigate and develop various approaches to help frontline health workers, policymakers, and populations to overcome the disease. While these techniques have been reviewed within individual disciplines, it is now timely to provide a cross-disciplinary overview of novel diagnostic and therapeutic approaches summarizing complementary efforts across multiple fields of research and technology. Accordingly, we reviewed and summarized various advanced novel approaches used for diagnosis and treatment of COVID-19 to help researchers across diverse disciplines on their prioritization of resources for research and development and to give them better a picture of the latest techniques. These include artificial intelligence, nano-based, CRISPR-based, and mass spectrometry technologies as well as neutralizing factors and traditional medicines. We also reviewed new approaches for vaccine development and developed a dashboard to provide frequent updates on the current and future approved vaccines

Development of a Clinical MALDI-ToF Mass Spectrometry Assay for SARS-CoV-2: Rational Design and Multi-Disciplinary Team Work.

The COVID-19 pandemic caused by the SARS-CoV-2 coronavirus has stretched national testing capacities to breaking points in almost all countries of the world. The need to rapidly screen vast numbers of a country's population in order to control the spread of the infection is paramount. However, the logistical requirement for reagent supply (and associated cost) of RT-PCR based testing (the current front-line test) have been hugely problematic. Mass spectrometry-based methods using swab and gargle samples have been reported with promise, but have not approached the task from a systematic analysis of the entire diagnostic process. Here, the pipeline from sample processing, the biological characteristics of the pathogen in human biofluid, the downstream bio- and physical-chemistry and the all-important data processing with clinical interpretation and reporting, are carefully compiled into a single high-throughput and reproducible rapid process. Utilizing MALDI-ToF mass spectrometric detection to viral envelope glycoproteins in a systems biology-multidisciplinary team approach, we have achieved a multifaceted clinical MALDI ToF MS screening test, primarily (but not limited to) SARS-CoV-2, with direct application to other future epidemics/pandemics that may arise. The clinical information generated not only includes SARS-CoV-2 coronavirus detection-(Spike protein fragments S1, S2b, S2a peaks), but other respiratory viral infections detected as well as an assessment of generalised oral upper respiratory immune response (elevated total Ig light chain peak) and a measure of the viral immune response (elevated intensity of IgA heavy chain peak). The advantages of the method include; (1) ease of sampling, (2) speed of analysis, and much reduced cost of testing. These features reveal the diagnostic utility of MALDI-ToF mass spectrometry as a powerful and economically attractive global solution

Saliva as a non-invasive diagnostic tool: COVID-19 and T2DM as case-study

Saliva has nowadays a vast research background of how it can mirror the body's health status. Specific salivary biomarkers have been already suggested for multiple diseases and is particularly useful for detecting infectious diseases. We focused on the importance of having a non-invasive, painless and self-collected fluid to study different aspects of two known pandemic diseases: COVID-19 and Type 2 Diabetes Mellitus. COVID-19 is the most impacting global pandemic of all time requiring frequent testing of populations. The necessity to identify cost-effective strategies for the detection of SARS-CoV-2 outbreak became a priority. Nasopharyngeal samples were considered the sampling golden standard but require a healthcare professional to collect the sample causing discomfort and pain to the patient. As saliva has proved successful in SARS-CoV-2 detection, a pooling strategy could be a good approach to decrease the number of individual tests and hazardous material waste which is also beneficial for the environment. We have tested this strategy on two hundred and seventy-nine saliva samples with pools of 10 and 20 randomized samples through RT-PCR. Cycle Threshold of the genes detected was 29.7. Consecutive reactions analysis of positive samples showed an equivalent cycle threshold average (p 0.05). We concluded that saliva-pool samples allowed effective SARS-CoV-2 screening on 10-sample and 20-sample pools. Our strategy was successfully applied in population-wide testing of more than 2000 individuals, showing that it is possible to use pooled saliva as diagnostic fluid for SARS-CoV-2 infection. The SARS-CoV-2 detection is well established with reliable methodologies including saliva as a detection fluid. In the opposite direction are the molecular alterations induced by this infectious virus which remain elusive. We developed a hybrid proteomics and in silico interactomics strategy to establish a COVID-19 salivary protein profile. The most distinctive proteins between healthy and COVID-19 samples were defined with the Partial Least-Squares Discriminant Analysis and the enrichment analysis was performed with FunRich software. In parallel, Protein-Protein virus-host interactome was identified with OralInt algorithm. Five dysregulated biological processes were identified in the COVID-19 proteome profile: Apoptosis, Energy Pathways, Immune Response, Protein Metabolism, and Transport. We identified 10 proteins (KLK 11, IMPA2, ANXA7, PLP2, IGLV2-11, IGHV3-43D, IGKV2-24, TMEM165, VSIG10, and PHB2) that had never been associated with SARS-CoV-2 infection, representing new evidence for the molecular profile behind COVID-19. Interactomics analysis showed viral influence on the host immune response, mainly through interaction with the degranulation of neutrophils. From our results, we can conclude that the virus also alters the host’s energy metabolism and interferes with apoptosis mechanisms. Type 2 Diabetes Mellitus is a chronic metabolic disease and is a major health risk due to its characteristic long-term complications. It is estimated that about 537 million people live with diabetes worldwide and will continue to increase. Diagnosis and glucose monitoring in diabetes are well established. However, monitoring the many of diabetes complications remains a challenge, compromising patients' prognosis and quality of life. We established a hybrid strategy that identified salivary markers of T2DM and its complications. From the functional analysis we highlight metabolic processes, response to stimulus, immune system processes and signalling as disrupted biological processes by the known relation with T2DM. The enrichment analysis identified 11 deregulated biological processes emphasizing 20 proteins directly related to complications in diabetes. Diabetic retinopathy, metabolic syndrome, insulin resistance, molecular impact of glucose and insulin homeostasis dysregulation, atherosclerosis, diabetic foot ulcer, protein catabolism and salivary gland function are diabetic complication capable of being monitored using saliva. We conclude that saliva has the potential to identify several molecules altered in diabetic patients compared to non-diabetic patients and that may be biomarkers not only of diabetes but also of the different complications of this disease. In conclusion, with this research we have confirmed the potential of saliva as a fluid of interest in both diagnosing and discovering new insights into diseases. Saliva was crucial in demonstrating the flexibility and capacity of its use demonstrated by the development of a reliable detection of SARS-CoV-2, the identification and discussion of molecular aspects of viral infection in and with the host, and the discovery of new markers in the diagnosis and monitoring of Type 2 Diabetes Mellitus.A Saliva tem atualmente um vasto background de investigação sobre como pode espelhar o estado de saúde do corpo. Já foram identificados biomarcadores salivares específicos para múltiplas doenças e são particularmente úteis para a monitorização de medicamentos. A saliva é um tipo de amostra particularmente favorável para a deteção de doenças infeciosas. Trata-se de um fluído não-invasivo, indolor e passível de auto recolha, ideal para estudar diferentes aspetos de duas doenças pandémicas conhecidas: COVID-19 e Diabetes Mellitus Tipo 2. A COVID-19 é uma das pandemias mais impactante de todos os tempos, exigindo testes frequentes às populações. A necessidade de identificar estratégias rentáveis para a deteção do surto de SARS-CoV-2 tornou-se uma prioridade global. As amostras nasofaríngeas foram consideradas como o padrão de amostragem, apesar de requerem um profissional de saúde qualificado para efetuar a colheita além de causar desconforto e dor ao doente. Uma vez que a saliva demonstrou ser bem-sucedida na deteção da SARS-CoV-2, o próximo passo seria criar uma estratégia de pools de amostras com vista a diminuir o número de testes individuais e de resíduos de materiais perigosos, o que também seria benéfico para o ambiente. Esta estratégia foi testada em duzentas e setenta e nove amostras de saliva com pools de 10 e 20 amostras aleatórias. O Cycle-Threshold médio dos genes detetados foi de 29,7. Reações de RT-PCT consecutivas em amostras positivas mostraram reprodutibilidade. A análise de comparação de amostras positivas individualizadas mostrou uma carga viral mediana mais elevada em amostras de saliva comparativamente a amostras nasofaríngeas. Concluímos que as amostras de saliva-pool permitiram um rastreio eficaz da SARS-CoV-2 em pools de 10 e de 20 amostras. Esta estratégia foi aplicada com sucesso em testes populacionais de mais de 2000 indivíduos, mostrando que é possível utilizar saliva em pool como líquido de diagnóstico para a infeção pelo SARS-CoV-2. Está demonstrado que a deteção de SARS-CoV-2 está bem estabelecida e com metodologias fiáveis, incluindo a saliva como fluido de deteção. No entanto as alterações moleculares induzidas por este vírus continuam por desvendar. Desenvolvemos uma estratégia proteómica híbrida para estabelecer um perfil proteico salivar COVID-19. As proteínas que mostram uma maior distinção entre amostras saudáveis e COVID-19 foram definidas através da análise PLS-DA e de enriquecimento funcional. Paralelamente, foram previstas as interações proteína-proteína entre o vírus e o hospedeiro. Foram identificados cinco processos biológicos desregulados no perfil do proteoma COVID-19: Apoptose, Vias de Energia, Resposta Imune, Metabolismo de Proteínas, e Transporte. Foram identificadas 10 proteínas (KLK 11, IMPA2, ANXA7, PLP2, IGLV2-11, IGHV3-43D, IGKV2-24, TMEM165, VSIG10 e PHB2) que nunca tinham sido associadas à infeção por SARS-CoV-2, representando novas provas do perfil molecular por detrás da COVID-19. A análise de interactómica mostrou influência do vírus na resposta imunitária do hospedeiro, principalmente através da interação com a desgranulação dos neutrófilos. A partir destes resultados, é possível concluir que o vírus altera não só a resposta imune, mas também o metabolismo energético do hospedeiro e interfere com os mecanismos de apoptose. A Diabetes Mellitus Tipo 2 é uma doença metabólica crónica e constitui um risco de saúde importante devido às suas complicações características a longo prazo. Estima-se que cerca de 537 milhões de pessoas vivem com diabetes em todo o mundo e continuarão a aumentar. O diagnóstico e a monitorização da glicose na diabetes estão atualmente bem estabelecidos. Contudo, a monitorização de muitas das complicações da diabetes continua a ser um desafio, comprometendo o prognóstico e a qualidade de vida dos pacientes. Tivemos como objetivo estabelecer uma estratégia híbrida que identificou os marcadores salivares do T2DM e as suas complicações. Da análise funcional destacamos processos metabólicos, resposta a estímulos, processos do sistema imunitário e sinalização como processos biológicos perturbados pela relação conhecida com o T2DM. A análise de enriquecimento identificou 11 processos biológicos desregulamentados em que demos destaque a 20 proteínas diretamente relacionadas com complicações na diabetes. A retinopatia diabética, síndrome metabólica, resistência à insulina, impacto molecular da glicose e desregulação da homeostase da insulina, aterosclerose, úlcera do pé diabético, catabolismo proteico e função da glândula salivar são complicações diabéticas capazes de serem monitorizadas usando saliva. Concluímos que a saliva tem o potencial de identificar proteínas alteradas em doentes diabéticos em comparação com não diabéticos e que podem ser biomarcadores não só da diabetes, mas também das diferentes complicações desta doença. Em conclusão, com esta investigação confirmamos o potencial da saliva como fluido de interesse tanto no diagnóstico como na descoberta de novos insights sobre doenças. A saliva foi crucial para demonstrar a flexibilidade e capacidade de seu uso demonstrada pelo desenvolvimento de uma deteção confiável de SARS-CoV-2, a identificação e discussão de aspetos moleculares da infeção viral no hospedeiro e com o hospedeiro, bem como a descoberta de novos marcadores no diagnóstico e monitorização da Diabetes Mellitus tipo 2

Aprotinin Inhibits SARS-CoV-2 Replication

Severe acute respiratory syndrome virus 2 (SARS-CoV-2) is the cause of the current coronavirus disease 19 (COVID-19) pandemic. Protease inhibitors are under consideration as virus entry inhibitors that prevent the cleavage of the coronavirus spike (S) protein by cellular proteases. Herein, we showed that the protease inhibitor aprotinin (but not the protease inhibitor SERPINA1/alpha-1 antitrypsin) inhibited SARS-CoV-2 replication in therapeutically achievable concentrations. An analysis of proteomics and translatome data indicated that SARS-CoV-2 replication is associated with a downregulation of host cell protease inhibitors. Hence, aprotinin may compensate for downregulated host cell proteases during later virus replication cycles. Aprotinin displayed anti-SARS-CoV-2 activity in different cell types (Caco2, Calu-3, and primary bronchial epithelial cell air–liquid interface cultures) and against four virus isolates. In conclusion, therapeutic aprotinin concentrations exert anti-SARS-CoV-2 activity. An approved aprotinin aerosol may have potential for the early local control of SARS-CoV-2 replication and the prevention of COVID-19 progression to a severe, systemic disease

Point-of-care diagnostics of covid-19: From current work to future perspectives

Coronaviruses have received global concern since 2003, when an outbreak caused by SARS‐CoV emerged in China. Later on, in 2012, the Middle‐East respiratory syndrome spread in Saudi Arabia, caused by MERS‐CoV. Currently, the global crisis is caused by the pandemic SARS‐ CoV‐2, which belongs to the same lineage of SARS‐CoV. In response to the urgent need of diagnostic tools, several lab‐based and biosensing techniques have been proposed so far. Five main areas have been individuated and discussed in terms of their strengths and weaknesses. The cell‐culture detection and the microneutralization tests are still considered highly reliable methods. The genetic screening, featuring the well‐established Real‐time polymerase chain reaction (RT‐PCR), represents the gold standard for virus detection in nasopharyngeal swabs. On the other side, immunoassays were developed, either by screening/antigen recognition of IgM/IgG or by detecting the whole virus, in blood and sera. Next, proteomic mass‐spectrometry (MS)‐based methodologies have also been proposed for the analysis of swab samples. Finally, virus-biosensing devices were efficiently designed. Both electrochemical immunosensors and eye‐based technologies have been described, showing detection times lower than 10 min after swab introduction. Alternative to swab‐based techniques, lateral flow point‐of‐care immunoassays are already commercially available for the analysis of blood samples. Such biosensing devices hold the advantage of being portable for on‐site testing in hospitals, airports, and hotspots, virtually without any sample treatment or complicated lab precautions

Absolute quantification of exogenous stimuli-induced nucleic acid modification dynamics with LC-MS

Modifications of nucleic acids comply different functions and are involved in genome or-ganization, cell differentiation, silencing, structure stability and enzyme recognition. Modi-fication abundances can be regulated intrinsically, like the incorporation of cap modifica-tions on viral RNA to evade the host immune response, but also extrinsically as a cause of damage, which can result in mutations or translational defects. Either way, modifications are highly dynamic. It is of great importance to trace and quantify these changes in order to understand the underlying mechanisms, which may offer a more divers applica-bility of RNA therapeutics and even facilitate the establishment of personalized medicine. Mass Spectrometry is a common technique to examine nucleic acids. However, mass spectrometry per se offers solely a static insight into the versatile dynamics of nucleic acid modifications. In order to circumvent this obstacle, Nucleic Acid Isotope Labeling coupled Mass Spectrometry (NAIL-MS) was developed. This powerful technique allows for absolute quantification on the one hand and on the other hand for examination of modification dynamics originating from endogenous or exogenous actuators. In 2010, the stress-induced reprogramming of tRNA modification in S. cerevisiae was reported. However, the underlying mechanisms remained to be elucidated. Few years later, the dynamics of RNA modifications and mechanisms like dilution, degradation and (de-)modification could be identified by the application of NAIL-MS. The first part of my dissertation deals with the examination of the extent of damage-induced alterations on nucleic acids. Therefore, a novel biosynthetically produced stable isotope labeled internal standard (SILIS) was established, to avoid the interference of signals with isotopologues generated in the stable isotope labeled pulse-chase experiments. Furthermore, the L-methionine-[2H3]-methyl labeling in S. cerevisiae was optimized to achieve full efficient labeling and thus again avoiding signal interferences with isotopologues due to inefficient labeling. Additionally, the tandem size exclusion chromatography was developed, allowing the time efficient purification of 28S/25S, 18S rRNA and tRNA in a single step. The appli-cation of improved stable isotope labeling and the facilitated purification of RNA popula-tions allowed for the examination of the stress-induced alterations in the RNA modifica-tion profile of S. cerevisiae. Thereby, the knowledge on stress-induced reprogramming of tRNA modifications in yeast could be expanded. Original and new transcripts could be discerned and in addition endogenous methylation could be differentiated from damage induced methylation. It was shown, that stress-induced alterations occur on original tRNA transcripts, whereas new transcripts were not affected. Moreover, the fast decrease of damage-induced methylations on 25S, 18S rRNA and tRNA in S. cerevisiae was demon-strated. Additionally, the formation of base damage on 2’-O-methylated nucleosides in rRNA upon methyl methanesulfonate (MMS) exposure were detected and thereby novel damage products of MMS could be identified. Furthermore, the application of NAIL-MS was expanded to study the endogenous and damage-induced methylome on the genomic levels in S. cerevisiae and E. coli. In parallel to the aforementioned findings, the fast dis-appearance of damage-induced methylations in the genome and transcriptome of S. cerevisiae and E. coli was shown. Apart from that, m7dG and m7G could be identified as the main damage products in the genome and transcriptome of both organisms. In parallel to prokaryotes and eukaryotes, the modifications in viral RNA are highly dy-namic. RNA viruses have high mutation rates and their modification abundances can vary during infection. So, several mutants and variants of the RNA virus SARS-CoV-2 emerged since 2019. It is necessary to understand the characteristics of the viral genome and the differences in mutants and variants in order to identify novel drug targets and optimize the application of available therapeutics and vaccines. Our previous work on absolute quantification of nucleic acid modifications in various organisms showed the strength of our LC-MS based approach. In the course of this study it was aimed at inves-tigating the viral RNA modification profile in the different mutants and variants of SARS-CoV-2. The absolute quantification of RNA modifications and the comparison to pub-lished reports lead to the assumption that observed modification densities are highly de-pendent on the cultivation and infection conditions as well as the purification method and verification of sample integrity is crucial for valid analysis. As outlined above, less is known about the genome of SARS-CoV-2 in terms of internal modifications. While the cap modification of the 5’ end of the SARS-CoV-2 genome is confirmed from many sides and is ascribed to regulate the host innate immune response and the viral replication. Hence, a better understanding of the viral capping mechanism is required in order to limit its contagiousness. Besides the interest in biological capping processes, the investigations on cap modifications become more relevant nowadays be-cause of mRNA therapeutics. The cap modification on engineered mRNA is necessary to prevent immunogenicity, improve intercellular stability and translation efficiency. Thus, therapeutic mRNA is engineered to resemble mature and processed eukaryotic mRNA, including the 5’ cap and the 3’ poly A tail. Currently, there are only a few published LC-MS methods for detection of cap modifications. Nevertheless, these methods include labor intensive sample preparation, long analyses times and have moderate sensitivity. In the course of my dissertation, the development and optimization of a time efficient and highly sensitive LC-MS method for absolute quantification of cap modifications is pre-sented. It includes an extensive method development, optimizing chromatographic and mass spectrometric parameters under consideration of short analysis time, low detection and quantification limits. For absolute quantification of cap modifications, an in vitro tran-scribed cap-SILIS was generated. Furthermore, limits of detection and quantification as well as the dynamic range for size and amount of macromolecules to be analyzed were determined. The high sensitivity allows for the analysis of RNA from synthetic but also from biological sources. The time efficiency is aspirational for ecologic and economic rea-sons, thus making this method suitable for high throughput analyses and industry. The identification and quantification of RNA modifications is getting more important with the significance of RNA therapeutics. In this work, efficient LC-MS based tools to study the extent of nucleic acid modifications are described. Insight into the stress-dependent regulation of the genome and transcriptome of common model organisms is given and a powerful method to quantify cap modifications is presented. These techniques can be used to study nucleic acid dynamics in clinical studies but also for quality control of RNA therapeutics

A combined approach of MALDI-TOF mass spectrometry and multivariate analysis as a potential tool for the detection of SARS-CoV-2 virus in nasopharyngeal swabs

Coronavirus disease 2019, known as COVID-19, is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The early, sensitive and specific detection of SARS-CoV-2 virus is widely recognized as the critical point in responding to the ongoing outbreak. Currently, the diagnosis is based on molecular real time RT-PCR techniques, although their implementation is being threatened due to the extraordinary demand for supplies worldwide. That is why the development of alternative and / or complementary tests becomes so relevant. Here, we exploit the potential of mass spectrometry technology combined with machine learning algorithms, for the detection of COVID-19 positive and negative protein profiles directly from nasopharyngeal swabs samples. According to the preliminary results obtained, accuracy =67.66 %, sensitivity =61.76 %, specificity =71.72 %, and although these parameters still need to be improved to be used as a screening technique, mass spectrometry- based methods coupled with multivariate analysis showed that it is an interesting tool that deserves to be explored as a complementary diagnostic approach due to the low cost and fast performance. However, further steps, such as the analysis of a large number of samples, should be taken in consideration to determine the applicability of the method developed.Fil: Rocca, María Florencia. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Red Nacional de Espectrometría de Masas Aplicada a la Microbiología Clínica; ArgentinaFil: Zintgraff, Jonathan Cristian. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud "Dr. C. G. Malbrán"; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Red Nacional de Espectrometría de Masas Aplicada a la Microbiología Clínica; ArgentinaFil: Dattero, María Elena. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; ArgentinaFil: Santos, Leonardo Silva. Universidad de Talca; ChileFil: Ledesma, Martin Manuel. Red Nacional de Espectrometría de Masas Aplicada A la Microbiología Clínica (renaem Argentina); Argentina. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vay, Carlos Alberto. Red Nacional de Espectrometría de Masas Aplicada a la Microbiología Clínica; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; ArgentinaFil: Prieto, Mónica Raquel. Red Nacional de Espectrometría de Masas Aplicada a la Microbiología Clínica; Argentina. Universidad de Buenos Aires; ArgentinaFil: Benedetti, Estefanía. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; ArgentinaFil: Avaro, Martín. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; ArgentinaFil: Russo, Mara Laura. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Nachtigall, Fabiane Manke. Universidad Autónoma de Chile; ChileFil: Baumeister, Elsa. Universidad Nacional de Santiago del Estero. Facultad de Humanidades Ciencias Sociales y de la Salud. Instituto de Estudios e Investigaciones en Enfermería; Argentina. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Instituto Nacional de Medicina Tropical; Argentin

Aptamers Targeting Membrane Proteins for Sensor and Diagnostic Applications.

Many biological processes (physiological or pathological) are relevant to membrane proteins (MPs), which account for almost 30% of the total of human proteins. As such, MPs can serve as predictive molecular biomarkers for disease diagnosis and prognosis. Indeed, cell surface MPs are an important class of attractive targets of the currently prescribed therapeutic drugs and diagnostic molecules used in disease detection. The oligonucleotides known as aptamers can be selected against a particular target with high affinity and selectivity by iterative rounds of in vitro library evolution, known as Systematic Evolution of Ligands by EXponential Enrichment (SELEX). As an alternative to antibodies, aptamers offer unique features like thermal stability, low-cost, reuse, ease of chemical modification, and compatibility with various detection techniques. Particularly, immobilized-aptamer sensing platforms have been under investigation for diagnostics and have demonstrated significant value compared to other analytical techniques. These "aptasensors" can be classified into several types based on their working principle, which are commonly electrochemical, optical, or mass-sensitive. In this review, we review the studies on aptamer-based MP-sensing technologies for diagnostic applications and have included new methodological variations undertaken in recent years