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Background & Aim: The pro-angiogenic, immunoregulatory and anti- inflammatory properties of MSCs are being exploited for the development of cellular therapies, including the treatment of graft versus host disease (GvHD), inflammatory bowel disease and COVID-19. SNBTS have developed a GMP process to bank umbilical cord MSCs (UC-MSCs) whereby we can reliably bank 100 vials of 10 million P2 UC-MSCs per cord. Each of these vials can be extensively expanded and stored for specific applications. The ultimate aim of the bank is for off-the-shelf clinical use, e.g., in GvHD or as an adjuvant therapy in Islet transplantations. Methods, Results & Conclusion(s): During process development, different basal media and supplements were screened for proliferation and MSC marker expression. Cells grown in promising media combinations were then tested for tri-lineage differentiation (identity), their chemokine/cytokine expression and T-cell inhibition (function) assessed. Medium selected for further GMP development and scale up was ultimately determined by all round performance and regulatory compliance. GMP-like UC-MSCs were shown to have immune-modulatory activity in T-cell proliferation assays at 4:1 or 16:1 ratios. Co-culture of UC-MSCs and freshly isolated leukocytes, +/- the immune activating agent LPS, show a dose dependent survival effect on leukocytes. In particular, neutrophils, which are normally very short lived in vitro demonstrated increased viability when co-cultured with UCMSCs. The survival effect was partially reproduced when UC-MSC were replaced with conditioned medium or cell lysate indicating the involvement of soluble factors. This improved neutrophil survival also correlates with results from leukocyte migration studies that demonstrate neutrophils to be the main cell type attracted to MSCs in in vitro and in vivo. Genetic modification of UC-MSC may improve their therapeutic potential. We have tested gene editing by CRISPR/Cas9 technology in primary UC-MSCS. The CXCL8 gene, highly expressed in UC-MSC, was targeted in isolates from several different donors with editing efficiencies of 78-96% observed. This translated to significant knockdown of CXCL8 protein levels in resting cells, however after stimulation levels of CXCL8 were found to be very similar in edited and non-edited UC-MSCs. This observation requires further study, but overall the results show the potential to generate future banks of primary UC-MSCS with genetically enhanced pro-angiogenic, immunoregulatory and/or anti-inflammatory activities.Copyright © 2023 International Society for Cell & Gene Therapy
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Objectives: The decision-making process for taking vaccination is influenced by a multitude of factors such as individual beliefs concerning vaccinations, trust in contextual forces, and sociodemographic. This study established a model to understand the relationship between people's beliefs in the safety, importance and effectiveness of vaccines, their trust in the medical advice from the government and doctors and their behaviors of having their children vaccinated from infectious diseases in low-and-middle-income countries (LMIC). Method(s): We structured a structural equation model with two latent variables, Motivation and Trust, and their relationships with the vaccination taking behavior. Motivation is constructed by people's beliefs in the safety, importance and effectiveness of vaccines and trust is constructed by people's trust in government, medical providers and scientists. This study used the 2018 Wellcome Global Monitor dataset and focused on people in 80 LMIC. The countries were divided into eight geographic regions: Eastern Africa, Central & Southern Africa, Norther Africa & Middle East, Western Africa, Central Asia, Southeast Asia, South Asia and Southern& Eastern Europe. Result(s): The latent variable Motivation is significantly positively associated with parental vaccination behaviors in all geographic areas except for South Asia and Western Africa. South Asia is the only area where the trust in government and medical system, providers had a significant association with vaccination behavior and such association is positive. Conclusion(s): In most LMIC, positive attitudes about vaccines are associated with an improved vaccine rate. Increasing people's belief in vaccines' importance, safety and effectiveness will be essential both for boosting vaccination rates and scaling up a vaccine for COVID-19. In South Asia, trust in the government and the public health system are important in deciding taking vaccines. In these countries, policymakers need to think of ways to improve people's trust in the public health system and further effectively communicate important health messages.Copyright © 2023
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Background/Objectives: The COVID-19 pandemic continues to threaten public health and burden healthcare systems worldwide. Whole SARS-CoV-2 genome sequencing has become essential for epidemiological monitoring and identification of new variants, which could represent a risk of increased transmissibility, virulence, or resistance to vaccines or treatment. In this study, we assess the performance of various target enrichment methods for whole SARS-CoV-2 sequencing. Method(s): We applied three target enrichment methods - two multiplex amplification methods and one hybridization capture - to the same set of nasopharyngeal patient samples (N = 93) in high-throughput mode. SARS-CoV-2 genome was obtained using short-read next-generation sequencing. Result(s): All three methods provided excellent breadth of coverage of SARS-CoV-2 genome (above 99%), albeit with vastly different sequencing depth (5-fold difference) and uniformity of coverage (20% difference in coefficient of variation). Poor local coverage has negative impact on variant calling in the concerned region, leading to an occasional allele drop-out (1.2% SNPs affected for one method). Conclusion(s): We discuss the performance of each target enrichment method and their potential for scaling up, in order to promote prospective programs of large-scale genomic surveillance of SARS-CoV-2 worldwide. Genomic surveillance will be crucial to overcoming the ongoing pandemic of COVID-19, despite its successive waves and continually emerging variants.
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While the World Health Organization strongly recommends HIV self-testing (HIVST) as an approach to HIV testing services, scale up has been minimal in low- and middle-income countries. Vietnam has successfully delivered HIVST at scale for key populations (KP), including men who have sex with men, transgender women, people who inject drugs, female sex workers and their partners. We reviewed data from the USAID/PATH Healthy Markets (2014-2021), including consumer surveys, HIVST usability and performance assessments, and service data to summarize the five stages of HIVST scale-up in Vietnam: 1) Assessing HIVST acceptability, preferences, and willingness to pay (WTP);2) Piloting HIVST;3) Developing HIVST policies and assessing products;4) Facilitating HIVST market authorization;and 5) Implementing differentiated service models. A '5A' approach was employed to shape HIVST markets, i.e., improving affordability, enhancing availability, assuring quality, ensuring appropriate design, and boosting awareness and demand. We assessed key factors related to HIVST supply and demand. In terms of supply, the median price people were willing to pay was US$4.3 per test. HIVST products with stringent regulatory approval successfully obtained free sale certificate registration, and blood-based HIVST products were highly accurate (99%-99.8%). Differentiated HIVST distribution models played a significant role in scaling-up HIVST and ensuring uninterrupted access to essential HIV services, e.g., pre-exposure prophylaxis monitoring during COVID-19 lock-downs. Related to demand and testing uptake, the majority of KPs accessing HIVST were first time HIV testers. Creative online-to-offline behavior change communication increased client awareness, trust, and use of HIVST. HIVST was successful in reaching first-time testers who may not otherwise test or seek facility-based care. HIVST is an effective strategy for reaching undiagnosed individuals and is accepted and preferred by KPs. HIVST scale-up requires enabling policy, intensive demand generation efforts, and differentiated service models.Copyright © 2023
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Background: The pandemic response measures have had significant global economic and health impacts with transient reductions in HIV clinic attendance and self-reported anti-retroviral therapy (ART) adherence reported in prior studies. Since viral suppression (VS) is an indication of ART adherence and effective service delivery, we assessed VS in the context of the COVID-19 pandemic in 3 African countries Methods: Since 2013, the African Cohort Study (AFRICOS) has enrolled individuals 18 years or older with and without HIV, in an approximate 5:1 ratio, at 12 clinics across 5 HIV care programs in Tanzania Uganda, Kenya, and Nigeria. For people living with HIV (PLWH), ART history was extracted from medical records and viral load was assessed at each visit. This assesses VS (< 1000 c/ml) before and during the COVID-19 pandemic (categorized into 4 surges and a consolidated non-surge period;defined in Table 1) among PLWH. Tanzania was excluded due to inadequate pandemic data. Logistic regression with generalized estimating equations, clustered by participant, was used to estimate odds ratios (ORs) and 95% confidence intervals (CI) comparing VS before and during COVID-19. Models are adjusted for age, sex, and program. Result(s): Of the 1741 study participants, 368 are from Uganda, 1156 are from Kenya, and 217 are from Nigeria;730 are males, 1011 are females, and 147 are under the age of 30. PLWH were less likely to be virally suppressed during the first surge period (OR 0.85, CI 0.46-1.56), but VS significantly increased during the second surge period (OR 1.95, CI 1.23-3.04) compared to the pre-COVID period. The third and fourth surge periods also saw a higher VS (table 1). Females are more likely to be virally suppressed than males (OR 1.58, CI 1.09- 2.29) and PLWH ages 40-49 have higher VS (OR 2.43, CI 1.32-4.48) compared to PLWH under. PLWH at the AFRICOS sites in Kenya and Nigeria show lower VS than the Ugandan cohort (ORs 0.46, CI 0.26-0.79 and OR 0.32, CI 0.17-0.60 respectively). Conclusion(s): The initial drop in VS may be attributed to reduced clinic access due to lockdowns. Many HIV programs supported by the President's Emergency Plan for AIDS Relief (PEPFAR) adapted their strategies to serve PLWH by scaling up community ART dispensing and multi-month dispensing (MMD) of ART for stable clients, which could have led to increased VS during the other surge periods.
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Background: COVID-19 testing is critical for identifying cases to prevent transmission. SARS-CoV-2 self-testing has the potential to increase diagnostic testing capacity and to expand access to hard-to-reach areas in low-andmiddle- income countries. We investigated the feasibility and acceptability of COVID-19 self-sampling and self-testing using SARS-CoV-2 Ag-Rapid Diagnostic Tests. Method(s): Between July 2021 to February 2022, we conducted a mixedmethods cross-sectional study examining self-sampling and self-testing using Standard Q and Panbio COVID-19 Ag Rapid Test Device in Urban and rural Blantyre, Malawi. Health care workers and adults (18y+) in the general population were systematically sampled. Result(s): Overall, 1,330 participants were enrolled of whom 674 (56.0%) were female with 664 for self-sampling and 666 for self-testing. Overall mean age was 30.7y (standard deviation [SD] 9.6). Self-sampling usability threshold for Standard Q was 273/333 (82.0%: 95% CI 77.4% to 86.0%) and 261/331 (78.8%: 95% CI 74.1% to 83.1%) for Panbio. Self-testing threshold was 276/335 (82.4%: 95% CI 77.9% to 86.3%) and 300/332 (90.4%: 95% CI 86.7% to 93.3%) for Standard Q and Panbio, respectively. Agreement between self-sample results and professional test results was 325/325 (100%) and 322/322 (100%) for Standard Q and Panbio, respectively. For self-testing, agreement was 332/333 (99.7%: 95% CI 98.3 to 100%) for Standard Q and 330/330 (100%: 95% CI 99.8 to 100%) for Panbio. Odds of achieving self-sampling threshold increased if the participant was recruited from an urban site (odds ratio [OR] 2.15 95% CI 1.44 to 3.23, P < .01. Compared to participants with primary school education those with secondary and those with tertiary achieved higher self-testing threshold OR 1.88 (95% CI 1.17 to 3.01), P = .01 and 4.05 (95% CI 1.20 to13.63), P = .02, respectively. Conclusion(s): One of the first studies to demonstrate high feasibility of self-testing using SARS-CoV-2 Ag-RDTs in low- and middle-income countries potentially supporting large scale-up.
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Background: Following acute COVID-19, some (~10-20%) individuals continue to experience a persistent variety of symptoms often referred to as long COVID- 19. However, evidence on long COVID-19 is limited from countries in Africa. We sought to describe the clinical presentation and factors associated with long COVID-19 in Zambia. Method(s): We conducted a cross-sectional analysis of routinely collected clinical information from patients receiving care in Zambia following SARS-CoV- 2 infection. Data were collected from 13 'post-acute COVID-19' (PAC-19) clinics established across Zambia to care for people following their acute infection. Long COVID-19 was defined as experiencing persistent symptoms >=4 weeks after initial diagnosis. Comorbidities detected at the time of SARS-CoV-2 infection were considered newly diagnosed. Severe illness was defined as acute COVID-19 that required supplemental oxygen therapy. We analyzed data from the first visit to the PAC-19 clinics and developed logistic regression models to assess factors associated with long COVID-19 at first visit to a PAC-19 clinic. Result(s): In total, 1,238 persons (< 1%) had >=1 visit to a PAC-19 clinic from August 2020 to April 2022 (out of ~319,500 confirmed cases in Zambia). Eight hundred twenty-three (66%) persons had been hospitalized for acute COVID-19 and the median length of stay was 8 days (interquartile range [IQR]: 4-16 days). Of these 1,238 persons, 641 (52%) were female while 403 (33%) had long COVID-19. The median age in persons with long COVID-19 was 54 years (IQR: 44-63) compared to 50 years (IQR: 37-61) in those without (p< 0.001). Cough (22%), fatigue (21%), and shortness of breath (15%) were frequently reported symptoms among persons with long COVID-19, while 4% had forgetfulness. Having severe illness (adjusted odds ratio [aOR] 2.8) and hospitalization for acute COVID-19 with length of stay >=15 days (aOR 13) were associated with having long COVID-19 (Table 1). Conclusion(s): Long COVID-19 was common among people attending PAC-19 clinics in Zambia, yet few persons with COVID-19 had attended a PAC-19 clinic. Those with severe illness were more likely to experience long COVID-19, which is consistent with other studies of long COVID-19. Given the burden of COVID-19 in Zambia, systems to care for patients with long COVID-19 might be needed in the future. Scaling up PAC-19 services and integrating into routine clinical care could improve access and further aid in understanding long COVID-19 in Zambia. (Table Presented).
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Background: People living with HIV (PLHIV) bear 20 times higher risk of acquiring tuberculosis (TB) compared to people without HIV. The World Health Organization recommends TB preventive treatment (TPT) for PLHIV to reduce this risk. However, according to the 2020 Global TB Report, only half of PLHIV were started on TPT globally in 2019, with the lowest coverage observed in low-income countries including Tanzania, where TPT provision is part of the standard of care for eligible PLHIV in Tanzania. We describe programmatic efforts to scale up TPT in 11 regions accounting for half of the 1.5 million PLHIV on ART in Tanzania. Method(s): Starting in 2018, PEPFAR, through the U.S. Centers for Disease Control and Prevention (CDC), supported the Government of Tanzania to accelerate TPT provision by: (1) training and mentoring healthcare workers, (2) integrating isoniazid into supply chain plans at the regional level, and (3) convening quarterly meetings at national and regional levels for program and supply chain monitoring and coordination. Additionally, CDC launched focused regional support interventions, with TPT among its priorities, aiming to facilitate real-time data-driven site monitoring, increased accountability, and on-the-ground coordination with local health authorities and implementing partners. We analyzed routine programmatic data reported in PEPFAR's data reporting system for fiscal years (FY) FY2018 through FY2021. Result(s): The number of PLHIV of all ages who initiated TPT increased from 67,510 in FY2018 to 268,909 in FY2019. Despite coinciding with the COVID-19 pandemic, the initiation numbers in FY2020 were sustained at 264,465 and dropped by about one-third in FY2021 (182,823) compared to the previous year. TPT completion rates among those initiated also showed a positive trend;38% in FY2018, 85% in FY2019, 90% in FY2020, and 91% in FY2021. Conclusion(s): Our findings demonstrate substantial acceleration of TPT initiation and a significant increase in TPT completion rates over the four-year period in 11 regions in Tanzania. The policy of once-in-a-lifetime TPT for PLHIV means fewer people are eligible for TPT over time, which might account for lower numbers of PLHIV initiated on TPT in FY2021. Completion remained high among those who initiated TPT. The strategic shift focusing on capacity building, supply chain strengthening, and site-level monitoring may have contributed to the improvements in TPT initiation and completion.
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Given the current coronavirus (COVID-19) situation around the world, we may have to face a long-term battle with coronavirus. It is necessary to prepare and stay resilient with some other techniques to improve air quality in buildings, especially in clinics and hospitals. In this paper, we have developed Ultraviolet-C (UVC) light-emitting diode (LED) modules which can be implemented in air ducts in heating, ventilation, and air conditioning system for airborne disinfection. An LED module is designed with LED panels as the basic unit so that it is easy to scale up to accommodate for air ducts with different sizes. Both experiments and simulations are carried out to study its disinfection performance. The results show that more than 76% and 85% of the pathogen can be inactivated within 60 and 90 min, respectively, in a meeting room with a volume of 107 m3 by using one LED module. Simulations for two LED modules show that the disinfection efficacy is more than two times compared to that of one LED module. In addition to the pathogen used in the experiments, the disinfection performance of the LED module for inactivation of SARS-CoV-2 virus based on the literature is investigated numerically. It shows that more than 99.70% of pathogens receive UV dose larger than 4.47 J/m2, leading to an almost 89.10% disinfection rate for SARS-CoV-2 virus within one hour using the two LED modules in the same meeting room. © 2023 Author(s).
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The study sought to explore the motivating factors for and barriers to the uptake of the coronavirus disease 2019 (COVID-19) vaccine in Zambia and recommend possible ways of addressing the challenges. The study used a qualitative case study approach, employing online media platforms as sources of data. Thematic analysis was to identify topics from posts, with each topic further grouped into themes. Motivating factors to vaccine uptake include an increase in reported cases of COVID-19, accessibility, and availability of vaccines, vaccine certificates required to travel, and credible sources of information. Barriers to uptake include lack of information, concerns about the safety of the vaccine, mistrust in Western medicine, lack of belief in conventional medicine versus belief in God, confusing information about eligibility, and belief that public figures used in the campaigns were not receiving the vaccine. Some of the strategies to encourage vaccination included the involvement of local scientists, demand generation through media platforms, community sensitization, vaccine materials, and creating an enabling environment for community involvement. The social media interactions on the COVID-19 vaccine revealed several factors that impede the uptake of the vaccine and encourage uptake. However, stakeholders' involvement in scaling up vaccination programs was reported to be one of the effective strategies to increase vaccine uptake in Zambia.Copyright © 2022 The Author(s). This open access article is distributed under a Creative Commons Attribution (CC-BY) 4.0 license.
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Over the past decade, compared to all other macromolecules lipid-based nanocarriers have proven to be an excellent carrier and delivery system for various pharmaceutical drugs of poor bioavailability. In addition to that, they exhibit exceptional qualities such as minimal toxicity, economical scale-up production, great biocompatibility, and high drug loading efficiency. In this study, we have discussed the various types of lipid nanoparticles, such as liposomes, nanostructured lipid carriers, solid lipid nanoparticles, and lipid polymer hybrid nanoparticles. We have also conferred in detail, the composition, shape and size, methods of preparation, advantages, and certain limitations associated with these lipid-based nanocarriers. Additionally, we have exclusively accounted for several examples of lipid-based nanomedicines that have either been approved and commercialized or are under the different phases of clinical trials. The current review overall focuses on the up-to-date research that has recently been published in view of developing lipid-based nanocarriers for various biological applications, including gene therapy, breast cancer therapy, and vaccine development.Copyright © 2022
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Background: The national multiple sclerosis (MS) registry is aimed at monitoring and improving quality of care and providing feedback to improve health outcomes by systematic recording of data. In 2018, the nationwide MS registry of Iran (NMSRI) was initiated for collecting epidemiological data and information on health care provision for patients with MS. The aim of the current study was to introduce the role of implementing coronavirus disease 2019 (COVID-19) scale-up registry protocol in NMSRI and arrange the national MS generality with information obtained during the COVID-19 pandemic. Method(s): The NMSRI group set up a program with crucial elements to collect the data of patients with MS who developed COVID-19. All MS cases with confirmed diagnosis of COVID-19 were enrolled in this study. New elements were considered to be added into the dataset, including demographic characteristics, definite diagnosis of COVID-19 and its symptoms, history of comorbidities, history of medications and hospitalization, changes in magnetic resonance imaging (MRI), and infection outcomes. Result(s): The COVID-19 data collection program was designed in NMSRI to collect data of MS cases with COVID-19 infection. The data collection protocol was explained to neurologists through an online training workshop. To the date of the study, 21 centers from 17 provinces of Iran were involved in the COVID-19 databases promoting NMSRI and 612 participants were registered successfully. Conclusion(s): We extended an agreement on data collection and developed it in NMSRI with various contributors to discover a critical need for COVID-19 awareness and monitor clinical training in MS.Copyright © 2021 Iranian Neurological Association, and Tehran University of Medical Sciences Published by Tehran University of Medical Sciences.
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In recent years, owing to the miniaturization of the fluidic environment, microfluidic technology offers unique opportunities for the implementation of nano drug delivery systems (NDDSs) production processes. Compared with traditional methods, microfluidics improves the controllability and uniformity of NDDSs. The fast mixing and laminar flow properties achieved in the microchannels can tune the physicochemical properties of NDDSs, including particle size, distribution and morphology, resulting in narrow particle size distribution and high drug-loading capacity. The success of lipid nanoparticles encapsulated mRNA vaccines against coronavirus disease 2019 by microfluidics also confirmed its feasibility for scaling up the preparation of NDDSs via parallelization or numbering-up. In this review, we provide a comprehensive summary of microfluidics-based NDDSs, including the fundamentals of microfluidics, microfluidic synthesis of NDDSs, and their industrialization. The challenges of microfluidics-based NDDSs in the current status and the prospects for future development are also discussed. We believe that this review will provide good guidance for microfluidics-based NDDSs.Copyright © 2023 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences
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Introduction: LenusCOPD patient and clinician apps include prompted daily patient-reported outcomes (PROs), self-management resources, structured clinical summary, wearable and home NIV data with asynchronous patientclinician messaging. To support patients with severe COPD during the COVID-19 pandemic we paused recruitment to the RECEIVER trial (NCT04240353) in March 2021 and provided LenusCOPD as routine care in the 'DYNAMICSCOT' service scale-up. Method(s): Data was censored on 31st August 2021 when 83 RECEIVER trial participants and 140 scale-up patients had completed minimum 1 year follow up. Outcomes including app usage, service workload, hospital admissions and mortality vs matched control cohort were determined from web analytic and de-identified EHR data. Result(s): Sustained patient app utilisation (avg 3.5 interactions/patient/week) was noted in both cohorts. Service management required 1 session of clinician time 5 days/week. There was a significant reduction in respiratory related admissions and occupied bed days comparing the 12 months pre- and post-onboarding and comparing RECEIVER & scale-up vs control patients. Figure 1 shows improved time to admission and death for the RECEIVER and scaleup cohorts of LenusCOPD users. PRO data showed stable quality of life scores across the cohorts. Conclusion(s): The DYNAMIC-SCOT project confirms the feasibility and benefits of scaling a digital service model for co-management of COPD.
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Background: Tailoring implementation strategies for scale-up involves engaging stakeholders, identifying implementation determinants, and designing implementation strategies to target those determinants. The purpose of this paper is to describe the multiphase process used to engage stakeholders in tailoring strategies to scale-up the Med-South Lifestyle Program, a research-supported lifestyle behavior change intervention that translates the Mediterranean dietary pattern for the southeastern US. Methods: Guided by Barker et al. framework, we tailored scale-up strategies over four-phases. In Phase 1, we engaged stakeholders from delivery systems that implement lifestyle interventions and from support systems that provide training and other support for statewide scale-up. In Phase 2, we partnered with delivery systems (community health centers and health departments) to design and pilot test implementation strategies (2014-2019). In Phase 3, we partnered with both delivery and support systems to tailor Phase 2 strategies for scale-up (2019-2021) and are now testing those tailored strategies in a type 3 hybrid study (2021-2023). This paper reports on the Phase 3 methods used to tailor implementation strategies for scale-up. To identify determinants of scale-up, we surveyed North Carolina delivery systems (n = 114 community health centers and health departments) and elicited input from delivery and support system stakeholders. We tailored strategies to address identified determinants by adapting the form of Phase 2 strategies while retaining their functions. We pilot tested strategies in three sites and collected data on intermediate, implementation, and effectiveness outcomes. Findings: Determinants of scale-up included limited staffing, competing priorities, and safety concerns during COVID-19, among others. Tailoring yielded two levels of implementation strategies. At the level of the delivery system, strategies included implementation teams, an implementation blueprint, and cyclical small tests of change. At the level of the support system, strategies included training, educational materials, quality monitoring, and technical assistance. Findings from the pilot study provide evidence for the implementation strategies' reach, acceptability, and feasibility, with mixed findings on fidelity. Strategies were only moderately successful at building delivery system capacity to implement Med-South. Conclusions: This paper describes the multiphase approach used to plan for Med-South scale-up, including the methods used to tailor two-levels of implementation strategies by identifying and targeting multilevel determinants.
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BACKGROUND: More than 3 y into the coronavirus 2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to undergo mutations. In this context, the Receptor Binding Domain (RBD) is the most antigenic region among the SARS-CoV-2 Spike protein and has emerged as a promising candidate for immunological development. We designed an IgG-based indirect enzyme-linked immunoassay (ELISA) kit based on recombinant RBD, which was produced from the laboratory to 10 L industry scales in Pichia pastoris. METHODS: A recombinant-RBD comprising 283 residues (31 kDa) was constructed after epitope analyses. The target gene was initially cloned into an Escherichia coli TOP10 genotype and transformed into Pichia pastoris CBS7435 muts for protein production. Production was scaled up in a 10 L fermenter after a 1 L shake-flask cultivation. The product was ultrafiltered and purified using ion-exchange chromatography. IgG-positive human sera for SARS-CoV-2 were employed by an ELISA test to evaluate the antigenicity and specific binding of the produced protein. RESULTS: Bioreactor cultivation yielded 4 g/l of the target protein after 160 h of fermentation, and ion-exchange chromatography indicated a purity > 95%. A human serum ELISA test was performed in 4 parts, and the ROC area under the curve (AUC) was > 0.96 for each part. The mean specificity and sensitivity of each part was 100% and 91.5%, respectively. CONCLUSION: A highly specific and sensitive IgG-based serologic kit was developed for improved diagnostic purposes in patients with COVID-19 after generating an RBD antigen in Pichia pastoris at laboratory and 10 L fermentation scales.
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COVID-19 , SARS-CoV-2 , Humans , COVID-19/diagnosis , Antibodies, Viral , Immunoglobulin GABSTRACT
The COVID-19 pandemic has reinforced Australia's need for diagnostic testing frameworks that are well-prepared, well-resourced, responsive, appropriately governed, scalable, interdisciplinary and collaborative.1 Point-of-care (POC) technologies offer diagnostic solutions capable of delivering individual, community and public health benefits in settings where: a) laboratory testing is not available, b) rapid turn-around time is needed, c) high loss to follow-up occurs in high-risk populations with standard of care cascades and/or d) disease transmission rates exceed laboratory response capacity. Key translational research derived from collaborative point-of-care testing networks for a) diabetes management (238 remote health services;3,233 operators;172,069 HbA1c and 51,379 urine albumin:creatinine ratio tests), b) acute care (106 remote health services;2,279 operators;32,950 blood gas, 32,689 cardiac troponin, 46,418 urea/electrolytes, 48,193 international normalised ratio tests), c) hepatitis C virus (HCV) (41 sites;110 operators;5,733 HCV tests;4,978 RNA, 755 antibody), d) syphilis screening (156 sites;1,412 operators), e) chlamydia, gonorrhea or trichomonas (51 sites;795 operators;>50,000 tests) or f) COVID-19 (101 remote health services, 733 operators, 72,624 tests) will be used to highlight operational, clinical, public health, and economic benefits of POC testing. Challenges associated with scale-up and accreditation pathways for decentralised POC testing will be discussed. Reference 1. Revised Testing Framework for COVID-19 in Australia, March 2022 Version 2.1. Communicable Disease Network Australia and Public Health Laboratory Network. Copyright © 2022
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In this article, Sara Dorman, Global Biopharma Market Manager at Roquette, discusses the benefits of single-use systems, including how they help accelerate a drug product's journey to market securely and more sustainably. Copyright © 2022, Frederick Furness Publishing. All rights reserved.
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World is facing a new pandemic called covid-19SARS-CoV-2) since a year ago. Unfortunately there is no treatment for Covid 19 nowadays as well as no potential therapies has been developed to overcome from coronavirus pandemic. Some potential drug molecules with combination have ability to respond for covid19 virus. From the research it was found that the reduction of viral load can be treated with hydroxychloroquine and azithromycin combination. We evaluate the mode of interactions of hydroxychloroquine and azithromycin with the dynamic site of SARS-CoV-2 coronavirus main protease. Molecular Structure-based computational approach viz. molecular docking simulations were performed to scale up their affinity and binding fitness of the docked complex of novel SARS-CoV-2 coronavirus protease and hydroxychloroquine and azithromycin. The natural inhibitor N3 of novel SARS-CoV-2 coronavirus protease were exhibited highest affinity in terms of MolDock score (-167.203Kcal/mol), and hydroxychloroquine was found with lowest target affinity (-55.917 Kcal/mol).The amino acid residue cysteine 145 and histidine 41 is bound covalently and formed hydrogen bond interaction with SARS-CoV-2 inhibitor known as inhibitor N3 as such, hydroxychloroquine and azithromycin also formed hydrogen bond interaction. The binding patterns of the inhibitor N3 of SARS-CoV-2 coronavirus main protease could be used as a guideline for medicinal chemist to explore their SARS-CoV-2 inhibitory potential. Copyright © RJPT All right reserved.