ARTICLE | doi:10.20944/preprints202208.0179.v1
Subject: Life Sciences, Other Keywords: In-house validation study; reproducibility precision; measurement uncertainty; prediction interval; uncertainty interval
Online: 9 August 2022 (10:56:40 CEST)
Measurement uncertainty is typically expressed in terms of a symmetric interval , where denotes the measurement result and the expanded uncertainty. However, in the case of heteroscedasticity, symmetric uncertainty intervals can be misleading. In this paper, a different approach for the calculation of uncertainty intervals is introduced. This approach is applicable when a validation study has been conducted with samples with known concentrations. It will be shown how, under certain circumstances, asymmetric uncertainty intervals arise quite naturally and lead to more reliable uncertainty intervals.
REVIEW | doi:10.20944/preprints201911.0176.v1
Online: 15 November 2019 (08:42:23 CET)
Infant cry is evolutionarily, psychologically, and clinically significant. During the last 60 years, several researchers and clinicians assessed the possibility of investigating the acoustical properties of cry for medical purposes. However, there is a lack of standardization in conducting and reporting cry-based studies. In this work, methodologies and procedures employed in infant cry analysis are reviewed, and best practices for reporting studies are provided. First, available literature on vocal and audio acoustic analysis have been examined to identify critical aspects of participant information, data collection, methods, and data analysis. Then, 180 peer-reviewed research articles have been assessed to certify the presence of identified critical information. Results show a general lack of critical description. Researchers in the field of infant cry need to agree on a standard set of criteria to report experimental studies, to better demonstrate the validity of the methods and obtained results.
REVIEW | doi:10.20944/preprints202006.0002.v1
Subject: Medicine & Pharmacology, Other Keywords: Biostatistics; Data management; Reproducibility; Workflow automation
Online: 2 June 2020 (09:24:25 CEST)
The complexity of analysis pipelines in biomedical sciences poses a severe challenge for the transparency and reproducibility of results. Researchers are increasingly incorporating software development technologies and methods into their analyses, but this is a quickly evolving landscape and teams may lack the capabilities to set up their own complex IT infrastructure to aid reproducibility. Basing a reproducible research strategy on readily available solutions with zero or low set-up costs whilst maintaining technological flexibility to incorporate domain-specific software tools is therefore of key importance. We outline a practical approach for robust reproducibility of analysis results. In our examples, we rely exclusively on established open-source tools and free services. Special emphasis is put on the integration of these tools with best practices from software development and free online services for the biostatistics domain.
CONCEPT PAPER | doi:10.20944/preprints202001.0176.v1
Subject: Keywords: community cyberinfrastructure; accessibility; reproducibility; interoperability; models
Online: 17 January 2020 (04:28:34 CET)
In an era of rapid global change, our ability to understand and predict Earth's natural systems is lagging behind our ability to monitor and measure changes in the biosphere. Bottlenecks in our ability to process information have reduced our capacity to fully exploit the growing volume and variety of data. Here, we take a critical look at the information infrastructure that connects modeling and measurement efforts, and propose a roadmap that accelerates production of new knowledge. We propose that community cyberinfrastructure tools can help mend the divisions between empirical research and modeling, and accelerate the pace of discovery. A new era of data-model integration requires investment in accessible, scalable, transparent tools that integrate the expertise of the whole community, not just a clique of ‘modelers’. This roadmap focuses on five key opportunities for community tools: the underlying backbone to community cyberinfrastructure; data ingest; calibration of models to data; model-data benchmarking; and data assimilation and ecological forecasting. This community-driven approach is key to meeting the pressing needs of science and society in the 21st century.
ARTICLE | doi:10.20944/preprints202004.0166.v1
Subject: Medicine & Pharmacology, General Medical Research Keywords: osteopathic manipulation; cranial osteopathy; reproducibility; osteopathic medicine
Online: 10 April 2020 (03:26:52 CEST)
Background and Objectives: The techniques directed to the cranial field in osteopathy are the most questioned due to the lack of scientific evidence. In osteopathic practice, manual palpation is essential and, therefore, measuring reliability is fundamental. The objective of study is to assess the reliability and validity of osteopathic treatment depending on experience. Materials and Methods: A cross-sectional study of reliability and validity was conducted. For measurements, a strain gauge was placed on the sphenobasilar synchondrosis of the skull base, and three maneuvers (lateral compression, anteroposterior compression and compression maneuver of the mastoids) were repeated 25 times each by osteopaths with different time of experience (5-10 years, 1-5 years, <1 year). Measurement averages were computed for each of the three maneuvers to verify the average effect of each group in comparison to that of the Gold Standard (GS) (>10 years of experience). Data were analyzed to check for inter- and intra-observer reliability using intra-class correlation coefficients (ICC). Results: Reliability and validity in 5-10 experience of observer 1 and observer 2 in the tree maneuvers was excellent (p<0.001) against GS. Poor or enough reproducibility and concordance were observed in osteopaths with less experience. Conclusion: Experience of osteopaths determines the efficacy of cranial maneuvers in osteopathic treatment for patients’ rehabilitation.
CONCEPT PAPER | doi:10.20944/preprints201901.0246.v2
Subject: Life Sciences, Endocrinology & Metabolomics Keywords: reproducibility; minimum guidelines; reporting; data analysis; reporting
Online: 8 March 2019 (09:06:02 CET)
Despite the proposal of minimum reporting guidelines for metabolomics over a decade ago, reporting on the data analysis step in metabolomics studies has been shown to be unclear and incomplete. Major omissions and a lack of logical flow render the data analysis’ sections in metabolomics studies impossible to follow, and therefore replicate or even imitate. Here, we propose possible reasons why the original reporting guidelines have had poor adherence and present an approach to improve their uptake. We present in this paper an R markdown reporting template file that guides the production of text and generates workflow diagrams based on user input. This R Markdown template contains, as an example in this instance, a set of minimum information requirements specifically for the data pre-treatment and data analysis section of biomarker discovery metabolomics studies, (gleaned directly from the original proposed guidelines by Goodacre at al.). These minimum requirements are presented in the format of a questionnaire checklist in an R markdown template file. The R Markdown reporting template proposed here can be presented as a starting point to encourage the data analysis section of a metabolomics manuscript to have a more logical presentation and to contain enough information to be understandable and reusable. The idea is that these guidelines would be open to user feedback, modification and updating by the metabolomics community via GitHub.
ARTICLE | doi:10.20944/preprints202007.0537.v1
Online: 23 July 2020 (08:10:38 CEST)
Energy use is of crucial importance for the global challenge of climate change but also an essential part of daily life. Hence, research on energy needs to be robust and valid. Other scientific disciplines have experienced a reproducibility crisis, that is, existing findings could not be reproduced in new studies, and energy research might be impacted as well. In this paper, we suggest the ‘TReQ’ approach to improve the research practices in the energy field and arrive at greater Transparency, Reproducibility, and Quality. We acknowledge the specific challenges of energy research and suggest a highly adaptable suite of tools that can be applied to research approaches across this multi-disciplinary and fast-changing field. In particular, we introduce preregistration of studies, making data and code publicly available, using preprints, and employing reporting guidelines to heighten the standard of research practices within the energy field. We argue that through wider adoption of these tools, we will be able to have greater trust in the findings of research used to inform evidence-based policy and practice in the energy field.
REVIEW | doi:10.20944/preprints202001.0378.v1
Subject: Life Sciences, Other Keywords: workflows; containers; cloud computing; Kubernetes; big data; reproducibility
Online: 31 January 2020 (05:15:01 CET)
Containers are gaining popularity in life science research as they provide a solution for encompassing dependencies of provisioned tools, simplify software installations for end users and offer a form of isolation between processes. Scientific workflows are ideal for chaining containers into data analysis pipelines to aid in creating reproducible analyses. In this manuscript we review a number of approaches to using containers as implemented in the workflow tools Nextflow, Galaxy, Pachyderm, Argo, Kubeflow, Luigi and SciPipe, when deployed in cloud environments. A particular focus is placed on the workflow tool’s interaction with the Kubernetes container orchestration framework.
ARTICLE | doi:10.20944/preprints201809.0543.v1
Subject: Medicine & Pharmacology, Behavioral Neuroscience Keywords: Open Science, Data Sharing, Neuroimaging, Reproducibility, Transparency, Reform
Online: 27 September 2018 (11:50:12 CEST)
Ongoing debates regarding the virtues and challenges of implementing open science for brain imaging research mirror those of the larger scientific community. The present commentary acknowledges the merits of arguments on both sides, as well as the underlying realities that have forced so many to feel the need to resist the implementation of an ideal. Potential sources of top-down reform are discussed, along with the factors that threaten to slow their progress. The potential roles of generational change and the individual are discussed, and a starter list of actionable steps that any researcher can take, big or small, is provided.
ARTICLE | doi:10.20944/preprints201810.0357.v1
Subject: Medicine & Pharmacology, Other Keywords: Reproducibility, quality, research integrity, universities, methods, science policy, rigor
Online: 16 October 2018 (11:33:46 CEST)
In recent years, biomedical research has faced increased scrutiny over issues related to reproducibility and quality in scientific findings(1-3). In response to this scrutiny, funding institutions and journals have implemented top-down policies for grant and manuscript review. While a positive step forward, the long-term merit of these policies is questionable given their emphasis on completing a check-list of items instead of a fundamental re-assessment of how scientific investigation is conducted. Moreover, the top-down style of management used to institute these policies can be argued as being ineffective in engaging the scientific workforce to act upon these issues. To meet current and future biomedical needs, new investigative methods that emphasize collective-thinking, teamwork, shared knowledge and cultivate change from the bottom-up are warranted. Here, a perspective on a new approach to biomedical investigation within the individual laboratory that emphasizes collaboration and quality is discussed.
ARTICLE | doi:10.20944/preprints202206.0320.v3
Subject: Life Sciences, Other Keywords: data; reproducibility; FAIR; data reuse; public data; big data; analysis
Online: 23 September 2022 (03:16:07 CEST)
With an increasing amount of "omics" data available publicly, there is a need for a guide on how to successfully download and use this data. The 10 simple rules for using public data are: 1) use public data in your research, 2) evaluate data for your use case, 3) check data reuse requirements and embargoes, 4) be aware of ethics for data reuse, 5) plan for data storage and compute requirements, 6) know what you are downloading, 7) download programmatically and verify integrity, 8) properly cite data, 9) make data FAIR and share, and 10) make pipelines and code FAIR and share. These rules are intended as a guide for researchers wanting to make use of available data and to increase data reuse and reproducibility.
ARTICLE | doi:10.20944/preprints202006.0295.v1
Subject: Medicine & Pharmacology, General Medical Research Keywords: insurance claims; reproducibility; propensity score; veridical data science; sensitivity analysis
Online: 24 June 2020 (09:51:17 CEST)
Medical insurance claims are becoming increasingly common data sources to answer a variety of questions in biomedical research. Although comprehensive in terms of longitudinal characterization of disease development and progression for a potentially large number of patients, population-based studies using these datasets require thoughtful modification to sample selection and analytic strategies, relative to other types of studies. Along with complex selection bias and missing data issues, claims-based studies are purely observational, which limits effective understanding and characterization of the treatment differences between groups being compared. All these issues contribute to a crisis in reproducibility and replication of comparative findings. This paper offers some practical guidance to the full analytical process, demonstrates methods for estimating causal treatment effects on several types of outcomes common to such studies, such as binary, count, time to event and longitudinally varying repeated measures outcomes, and aims to increase transparency and reproducibility. We provide an online version of the paper with readily implementable code for the entire analysis pipeline to serve as a guided tutorial for practitioners. The online version can be accessed at https://rydaro.github.io/. The analytic pipeline is illustrated using a sub-cohort of patients with advanced prostate cancer from the large Clinformatics TM Data Mart Database (OptumInsight, Eden Prairie, Minnesota), consisting of 73 million distinct private payer insurees from 2001-2016.
SHORT NOTE | doi:10.20944/preprints202001.0196.v1
Subject: Biology, Entomology Keywords: reproducibility; open access; data curation; data mangement; pre-print servers
Online: 18 January 2020 (09:05:49 CET)
The ability to replicate scientific experiments is a cornerstone of the scientific method. Sharing ideas, workflows, data, and protocols facilitates testing the generalizability of results, increases the speed that science progresses, and enhances quality control of published work. Fields of science such as medicine, the social sciences, and the physical sciences have embraced practices designed to increase replicability. Granting agencies, for example, may require data management plans and journals may require data and code availability statements along with the deposition of data and code in publicly available repositories. While many tools commonly used in replicable workflows such as distributed version control systems (e.g. “git”) or scripted programming languages for data cleaning and analysis may have a steep learning curve, their adoption can increase individual efficiency and facilitate collaborations both within entomology and across disciplines. The open science movement is developing within the discipline of entomology, but practitioners of these concepts or those desiring to work more collaboratively across disciplines may be unsure where or how to embrace these initiatives. This article is meant to introduce some of the tools entomologists can incorporate into their workflows to increase the replicability and openness of their work. We describe these tools and others, recommend additional resources for learning more about these tools, and discuss the benefits to both individuals and the scientific community and potential drawbacks associated with implementing a replicable workflow.
ARTICLE | doi:10.20944/preprints201909.0122.v1
Subject: Medicine & Pharmacology, Other Keywords: open health; simple rules; ethics; reproducibility; research significance; open science
Online: 11 September 2019 (13:27:26 CEST)
We are witnessing a dramatic transformation in the way we do science. In recent years, significant flaws with existing scientific methods have come to light, including lack of transparency, insufficient involvement of stakeholders, disconnection from the public, and limited reproducibility of research findings. These concerns have sparked a global movement to revolutionize scientific practice and the emergence of Open Science. This new approach to science extends principles of openness to the entire research cycle, from hypothesis generation to data collection, analysis, replication, and translation from research to practice. Open Science seeks to remove all barriers to conducting high quality, rigorous, and impactful scientific research by ensuring that the data, methods, and opportunities for collaboration are open to all. Emerging digital technologies and "big data" (see "Ten simple rules for responsible big data research") have further accelerated the Open Science movement by affording new approaches to data sharing, connecting researcher networks, and facilitating the dissemination of research findings. Open scientific practices are also having a profound impact on the health sciences and medical research, and specifically how we conduct clinical research with human participants. Human health research necessitates careful considerations for practicing science in an ethical manner. There is also a particular urgency to human health research since the goal is to help people, so doing good science takes on a different meaning than simply doing science well. It also implores the scientist to reassess the conventional view of human health research as a pursuit conducted by scientists on human subjects, and lays a greater emphasis on inclusive and ethical practices to ensure that the research takes into account the interests of those who would be most impacted by the research. Openness in the context of human health research also raises greater concerns about privacy and security and presents more opportunities for people, including participants of research studies, to contribute in every capacity. At the core of open health research, scientific discoveries are not only the product of collaboration across disciplines, but must also be owned by the community that is inclusive of researchers, health workers, and patients and their families. To guide successful open health research practices, it is essential to carefully consider and delineate its guiding principles. This editorial is aimed at individuals participating in health science in any capacity, including but not limited to people living with medical conditions, health professionals, study participants, and researchers spanning all types of disciplines. We present ten simple rules that, while not comprehensive, offer guidance for conducting health research with human participants in an open, ethical, and rigorous manner. These rules can be difficult, resource-intensive, and can conflict with one another. They are aspirational and are intended to accelerate and improve the quality of human health research. Work that fails to follow these rules is not necessarily an indication of poor quality research, especially if the reasons for breaking the rules are considered and articulated (see rule 6: document everything). While most of the responsibility of following these rules falls on researchers, anyone involved in human health research in any capacity can apply them.
ARTICLE | doi:10.20944/preprints201804.0068.v1
Subject: Keywords: reproducibility crisis; replication crisis; data reliability; bias; publication bias; meta-research
Online: 5 April 2018 (10:54:09 CEST)
A lack of data reproducibility (“reproducibility crisis”) is debated across many scientific and medical disciplines. A systematic analysis of the available evidence on the reliability of scientific data revealed that, although the existence of a reproducibility crisis is widely perceived, conclusive data on the scale of the problem are lacking. Most importantly we found that, although the debate is largely focused on methodological issues, researcher conduct defines research standards and in turn data reliability. The availability of reliable methods itself does not guarantee good practice. Moreover, research is typically characterised by a lack of established methods due to its immanent novelty. Despite the crucial importance of researcher conduct, research and conclusive data on the determinants of researcher behaviour are missing. In conclusion, meta-research is urgently needed that establishes an understanding of the factors that determine researcher behaviour. This knowledge can then be used to implement and iteratively improve measures, which incentivise researchers to apply the highest standards resulting in high quality data.
ARTICLE | doi:10.20944/preprints202011.0064.v1
Subject: Medicine & Pharmacology, Allergology Keywords: manual muscle testing; neuromuscular diagnostics; force profiles; reproducibility; Adaptive Force; handheld device
Online: 2 November 2020 (16:36:04 CET)
The manual muscle test (MMT) is a flexible diagnostic tool, which is used in many disciplines, applied in several ways. The main problem is the subjectivity of the test. The MMT in the version of a “break test” depends on the tester’s force rise and the patient’s ability to resist the applied force. As a first step, the investigation of the reproducibility of the testers’ force profiles is required for valid application. The study examined the force profiles of n=29 testers (n=9 experiences (Exp), n=8 little experienced (LitExp), n =12 beginners (Beg)). The testers performed 10 MMTs according to the test of hip flexors, but against a fixed leg to exclude the patient’s reaction. A handheld device recorded the temporal course of the applied force. The results show significant differences between Exp and Beg concerning the starting force (padj=0.029), the ratio of starting to maximum force (padj=0.005) and the normalized mean Euclidean distances between the 10 trials (padj=0.015). The slope is significantly higher in Exp vs. LitExp (p=0.006) and Beg (p=0.005). The results also indicate that experienced testers show inter-tester differences and partly even a low intra-tester reproducibility. That highlights the necessity of an objective MMT-assessment. Furthermore, an agreement on a standardized force profile is required – a suggestion is given.
ARTICLE | doi:10.20944/preprints202008.0664.v1
Subject: Engineering, Other Keywords: Microbial electrolysis cells; Linear sweep voltammetry; Counter electrode; Coefficient of variation; Reproducibility
Online: 30 August 2020 (11:45:33 CEST)
Electrode is a key component in a microbial electrolysis cell (MEC) and it needs significant improvement for practical implementation of MEC. For effective development of electrode technology, accurate and reproducible analytical methods are very important. Linear sweep voltammetry (LSV) is an essential analytical method for evaluating electrode performance; however, it has not been firmly established yet in the MEC field. In this study, biological brush (BB), abiotic brush (AB), Pt wire (PtW), stainless steel wire (SSW) and mesh (SSM)) were tested to explore the most suitable counter electrode in different medium conditions. Coefficient of variation (CV) for Imax of LSV were comparatively analyzed. In BB-anode LSV, SSW (0.48%) and SSM (2.17%) showed higher reproducibility as a counter electrode. In SSM-cathode LSV, BB (1.76%) and PtW (2.01%) produced more reproducible results. In the Ni-AC-SSM-cathode LSV, PtW (3.54%) and BB (8.81%) produced more reproducible result. It shows electrode used in the operation is an appropriate counter electrode in the acetate-added condition. However, in the absence of acetate, PtW (1.24%) and BB (1.71%) produced more reproducible results in SSM cathode and PtW (0.61%) and SSW (1.21%) did in the Ni-AC-SSM-cathode, showing PtW is an appropriate counter electrode. These results also shows that PtW is an appropriate counter electrode in cathode LSV.
ARTICLE | doi:10.20944/preprints202107.0134.v2
Subject: Medicine & Pharmacology, Clinical Neurology Keywords: Diffusion Magnetic Resonance Imaging; White Matter; Fractional anisotropy; Multi-centre; Reproducibility; Imaging artefacts; Ageing
Online: 6 September 2021 (13:20:18 CEST)
In clinical diagnostics and longitudinal studies, the reproducibility of MRI assessments is of high importance in order to detect pathological changes, but developments in MRI hard- and software often outrun extended periods of data acquisition and analysis. This could potentially introduce artefactual changes or mask pathological alterations. However, if and how changes of MRI hardware, scanning protocols or preprocessing software affect complex neuroimaging outcomes from e.g. diffusion weighted imaging (DWI) remains largely understudied. We therefore compared DWI outcomes and artefact severity of 121 healthy participants (age range 19-54 years) who underwent two matched DWI protocols (Siemens product and Center for Magnetic Resonance Research sequence) at two sites (Siemens 3T Magnetom Verio and Skyrafit). After different preprocessing steps, fractional anisotropy (FA) and mean diffusivity (MD) maps,obtained by tensor fitting, were processed with tract-based spatial statistics (TBSS). Inter-scanner and inter-sequence variability of skeletonised FA values reached up to 5% and differed largely in magnitude and direction across the brain. Skeletonised MD values differed up to 14% between scanners. We here demonstrate that DTI outcome measures strongly depend on imaging site and software, and that these biases vary between brain regions. These regionally inhomogeneous biases may exceed and considerably confound physiological effects such as ageing, highlighting the need to harmonise data acquisition and analysis. Future studies thus need to implement novel strategies to augment neuroimaging data reliability and replicability.
ARTICLE | doi:10.20944/preprints201905.0031.v1
Subject: Life Sciences, Biophysics Keywords: functional networks; functional magnetic resonance imaging; connectome; connectivity matrices; graphs; reproducibility; granger causality; transfer entropy
Online: 6 May 2019 (07:47:11 CEST)
A growing number of studies focus on methods to estimate and analyze the functional connectome of the human brain. Graph theoretical measures are commonly employed to interpret and synthesize complex network-related information. While resting state functional MRI (rsfMRI) is often employed in this context, is known to exhibit poor reproducibility, a key factor which is commonly neglected in typical cohort studies using connectomics-related measures as biomarkers. We aimed to fill this gap by analyzing and comparing inter- and intra- subject variability of connectivity matrices as well as graph-theoretical measures in a large (n=1003) database of young healthy subjects which underwent four consecutive rsfMRI sessions. We analyzed both directed (Granger Causality and Transfer Entropy) and undirected (Pearson Correlation and Partial Correlation) time-series association measures and related global and local graph-theoretical measures. While matrix weights exhibit a higher reproducibility in undirected as opposed to directed methods, this difference disappears when looking at global graph metrics and, in turn, exhibits strong regional dependence in local graphs metrics. Our results warrant caution in the interpretation of connectivity studies, and serve as a benchmark for future investigations by providing quantitative estimates for the inter- and intra- subject variabilities in both directed and undirected connectomic measures.
Subject: Physical Sciences, Optics Keywords: surface-enhanced fluorescence; quenching; Rhodamine 6G; hot spot; separation layer; high reproducibility; finite difference time domain
Online: 22 December 2019 (13:45:00 CET)
The surface enhanced fluorescence（SEF）detection bases by plasmonic nanopillars array with nanoparticles has opened up a new gate in the application of biological imaging and sensing. The fluorescence enhancement of the probe molecule depends on its position in equilibrium, which is close to the hot spot leading to the electromagnetic field enhancement, but not too close to the metal surface resulting in quenching. Here, a large scale SiO2-Ag-cicada wing SEF substrate was fabricated by magnetron sputtering with correction enhancement factor of 797.6. Thereinto the cicada wing provides the skeleton of the nanopillars array structure, the deposited Ag constructs two kinds of hot spots, and SiO2 forms a separation layer to prevent quenching. Moreover, the substrate exhibited good reproducibility, high sensitivity with low limits of detection (LOD) and high stability for oxidation resistance. We propose that SEF substrate with modification of SiO2 can not only improve the enhancement performance, but also expanding its application in the biological investigations.
Subject: Medicine & Pharmacology, Other Keywords: Reproducibility, Mathematical Modeling, Multiscale Modeling, Translational Research, Biomedical Research, Experimental Biology, Clinical Research Article Type: Essay
Online: 23 May 2018 (16:18:52 CEST)
The “Crisis of Reproducibility” has received considerable attention both within the scientific community and without. While factors associated with scientific culture and practical practice are most often invoked, I propose that the Crisis of Reproducibility is ultimately a failure of generalization with a fundamental scientific basis in the methods used for biomedical research. The Denominator Problem describes how limitations intrinsic to the two primary approaches of biomedical research, clinical studies and pre-clinical experimental biology, lead to an inability to effectively characterize the full extent of biological heterogeneity, which compromises the task of generalizing acquired knowledge. Drawing on the example of the unifying role of theory in the physical sciences, I propose that multi-scale mathematical and dynamic computational models, when mapped to the modular structure of biological systems, can serve a unifying role as formal representations of what is conserved and similar from one biological context to another. This ability to explicitly describe the generation of heterogeneity from similarity addresses the Denominator Problem and provides a scientific response to the Crisis of Reproducibility.
ARTICLE | doi:10.20944/preprints202203.0229.v1
Subject: Life Sciences, Biochemistry Keywords: SARS-CoV-2 antibody; reproducibility crisis; peptide mass fingerprinting; monoclonal antibody; trace-ability; identity; antibody identification; antibody light chain; MALDI-TOF-MS
Online: 16 March 2022 (10:01:41 CET)
During the SARS-CoV-2 pandemic, many virus-binding monoclonal antibodies have been developed for clinical and diagnostic purposes. This underlines the importance of antibodies as universal bioanalytical reagents. However, little attention is given to the reproducibility crisis that scientific studies are still facing to date. In a recent study, not even half of all research antibodies mentioned in publications could be identified at all. This should spark more efforts in the search for practical solutions for the traceability of antibodies. For this purpose, we used thirty-five monoclonal antibodies against SARS-CoV-2 to demonstrate how sequence-independent antibody identification can be achieved by simple means applied onto the protein. First, we examined the intact and light chain masses of the antibodies relative to the reference material NIST-mAb 8671. Already half of the antibodies could be identified based solely on these two parameters. In addition, we developed two complementary peptide mass fingerprinting methods with MALDI-TOF-MS that can be performed in 45 minutes and had a combined sequence coverage of over 80%. One method is based on the partial acidic hydrolysis of the protein by 5 mM of sulfuric acid at 99 °C. Furthermore, we established a fast way for a tryptic digest without an alkylation step. We were able to show that the distinction of clones is possible simply by a brief visual comparison of the mass spectra. In this work, two clones originating from the same immunization gave the same fingerprints. Later, a hybridoma sequencing confirmed the sequence identity of these sister clones. In order to automate the spectral comparison for larger libraries of antibodies, we developed the online software ABID 2.0 (https://gets.shinyapps.io/ABID/). This open-source software determines the number of matching peptides in the fingerprint spectra. We propose that publications and other documents critically relying on monoclonal antibodies with unknown amino acid sequences should include at least one antibody fingerprint. By fingerprinting an antibody in question, its identity can be confirmed by comparison with a library spectrum at any time and context.
ARTICLE | doi:10.20944/preprints202002.0207.v1
Subject: Life Sciences, Biotechnology Keywords: Antibody ID; antibody registry; Research Resource Identifier; RRID; reproducibility; quality control; documentation; traceability; clones; biochemical reagents; diagnostics; immunoassays; ELISA; western blot; immunohistochemistry; microarray; biosensor
Online: 15 February 2020 (15:46:27 CET)
Thousands of antibodies for diagnostic and other analytical purposes are on the market. However, it is often difficult to identify duplicates, reagent changes, and to assign the correct original publications to an antibody. This slows down scientific progress and might even be a cause of irreproducible research and a waste of resources. Recently, activities were started to suggest the sole use of recombinant antibodies in combination with the open communication of their sequence. In this case, such uncertainties should be eliminated. Unfortunately, this approach seems to be rather a long-term vision since the development and manufacturing of recombinant antibodies remain quite expensive in the foreseeable future. Also, nearly all commercial antibody suppliers may be reluctant to publish the sequence of their antibodies, since they fear counterfeiting. De-novo sequencing of antibodies is also not feasible today for a reagent user without access to the hybridoma clone. Nevertheless, it seems to be crucial for any scientist to have the opportunity to identify an antibody undoubtedly to guarantee the traceability of any research activity using antibodies from a third party as a tool. For this purpose, we developed a method for the identification of antibodies based on a MALDI-TOF-MS fingerprint. To circumvent lengthy denaturation, reduction, alkylation, and enzymatic digestion steps, the fragmentation was performed with a simple formic acid hydrolysis step. Eighty-nine unknown monoclonal antibodies were used for this study to examine the feasibility of this approach. Although the molecular assignment of peaks was rarely possible, antibodies could be easily recognized in a blinded test, simply from their mass-spectral fingerprint. A general protocol is given, which could be used without any optimization to generate fingerprints for a database. We want to propose that in most scientific projects relying critically on antibody reagents, such a fingerprint should be established to prove and document the identity of the used antibodies and to assign a specific reagent to a datasheet of a commercial supplier, a public database record or an antibody ID.
ARTICLE | doi:10.20944/preprints202205.0223.v1
Subject: Engineering, Other Keywords: digitization; virtualization; digital twin; blockchain; crowdsourcing; decentralization; non-fungible token; NFT; smart contract; oracle; tokenization; digital ownership; consensus; governance; trust; incentivization; staking; reputation systems; reproducibility crisis; exponentiality; digital twin; metaverse; DeSci; decentralized science; citizen science; open science; distributed ledger; digital scarcity
Online: 17 May 2022 (05:50:03 CEST)
Fundamental science and applied research and technology development (RTD) are facing significant challenges that particularly compound to the notorious credibility, reproducibility, funding and sustainability crises. The underlying, serious shortcomings are substantially amplified by a metrics-obsessed publication culture, and a growing cohort of academics fishing for fairly stagnant (public) funding budgets. This work presents, for the first time, a groundbreaking strategy to successfully address these severe issues; the novel strategy proposed here leverages the distributed ledger technology (DLT) “blockchain” to capitalize on cryptoeconomic mechanisms, such as tokenization, consensus, crowdsourcing, smart contracts, reputation systems as well as staking, reward and slashing mechanisms. This powerful toolbox, which is so far widely unfamiliar to traditional scientific and RTD communities (“TradSci”), is synergistically combined with the exponentially growing computing capabilities for virtualizing experiments through digital twin methods in a future scientific “metaverse”. Project contributions, such as hypotheses, methods, experimental data, modelling, simulation, assessment, predictions and directions are crowdsourced using blockchain, and captured by so-called non-fungible tokens (“NFTs”). The so enabled, highly integrative approach, termed decentralized science (“DeSci”), is destined to move research out of its present silos, and to markedly enhance quality, credibility, efficiency, transparency, inclusiveness, sustainability, impact, and sustainability of a wide spectrum of academic and commercial research initiatives.