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Protein biomarkers in biological fluids represent an important resource for improving the clinical management of diseases. Current proteomics technologies are capable of performing high-throughput and multiplex profiling in different types of fluids, often leading to the shortlisting of tens of candidate biomarkers per study. However, before reaching any clinical setting, these discoveries require thorough validation and an assay that would be suitable for routine analyses. In the path from biomarker discovery to validation, the performance of the assay implemented for the intended protein quantification is extremely critical toward achieving reliable and reproducible results. Development of robust sandwich immunoassays for individual candidates is challenging and labor and resource intensive, and multiplies when evaluating a panel of interesting candidates at the same time. Here we describe a versatile pipeline that facilitates the systematic and parallel development of multiple sandwich immunoassays using a bead-based technology.A novel protein microarray technology, called high-density nucleic acid programmable protein array (HD-NAPPA), enables the serological screening of thousands of proteins at one time. buy SCH66336 HD-NAPPA extends the capabilities of NAPPA, which produces protein microarrays on a conventional glass microscope slide. By comparison, HD-NAPPA displays proteins in over 10,000 nanowells etched in a silicon slide. Proteins on HD-NAPPA are expressed in the individual isolated nanowells, via in vitro transcription and translation (IVTT), without any diffusion during incubation. Here we describe the method for antibody biomarker identification using HD-NAPPA, including four main steps (1) HD-NAPPA array protein expression, (2) primary antibodies (serum/plasma) probing, (3) secondary antibody visualization, and (4) image scanning and data processing.Chronic diseases are the leading cause of disability and responsible for about 63% of deaths worldwide. Among the noninfectious chronic diseases with the highest incidence are cancer and neurodegenerative diseases. Although they have been extensively studied in the last years, there is still an urgent need to find and elucidate the molecular mechanisms underlying their formation and progression to get an early diagnosis and find new therapeutic targets of intervention. Beyond other microarray-based proteomic techniques more extensively used because of their commercial availability, such as protein and antibody microarrays, phage microarrays are another kind of protein microarrays useful for the identification and characterization of disease-specific humoral immune responses and to get further insights into these devastating diseases. Here, we describe the integration and utilization of phage microarrays, which offer such a combination of sensitivity and cost-effective multiplexing capabilities that makes them an affordable strategy for the characterization of humoral immune responses in multiple diseases.The systematic design and construction of customized protein microarrays are critical for the further successful screening of biological samples in biomedical research projects. In general protein microarrays are classified according to the content, detection method, and printing methodology, among others. Here, we are focused on the type of printing contact and noncontact. Both approaches have advantages and disadvantages; however, in any of the approaches, a prior well design and systematic preparation of materials and/or instruments required for the customized antibody arrays is critical. In this chapter, the process for an antibody microarray by a noncontact printer is described in detail from the preparation of array content to the analysis, including quality control steps.As we approach the twentieth anniversary of completing the international Human Genome Project, the next (and arguably most significant) frontier in biology consists of functionally understanding the proteins, which are encoded by the genome and play a crucial role in all of biology and medicine. To accomplish this challenge, different proteomics strategies must be devised to examine the activities of gene products (proteins) at scale. Among them, protein microarrays have been used to accomplish a wide variety of investigations such as examining the binding of proteins and proteoforms to DNA, small molecules, and other proteins; characterizing humoral immune responses in health and disease; evaluating allergenic proteins; and profiling protein patterns as candidate disease-specific biomarkers. In Protein Microarray for Disease Analysis Methods and Protocols, expert researchers involved in the field of protein microarrays provide concise descriptions of the methodologies that they currently use to fabricate microarrays and how they apply them to analyze protein interactions and responses of proteins to dissect human disease.Biological diversity is the basis for, and an indicator of biosphere integrity. Together with climate change, its loss is one of the two most important planetary boundaries. A halt in biodiversity loss is one of the UN Sustainable Development Goals. Current changes in biodiversity in the vast landmass of Siberia are at an initial stage of inventory, even though the Siberian environment is experiencing rapid climate change, weather extremes and transformation of land use and management. Biodiversity changes affect traditional land use by Indigenous People and multiple ecosystem services with implications for local and national economies. Here we review and analyse a large number of scientific publications, which are little known outside Russia, and we provide insights into Siberian biodiversity issues for the wider international research community. Case studies are presented on biodiversity changes for insect pests, fish, amphibians and reptiles, birds, mammals and steppe vegetation, and we discuss their causes and consequences.Zoonotic disease emergence has become a core concern of biodiversity conservation amid the ongoing impacts of the COVID-19 pandemic. Major international conservation groups now comprehensively center larger human-nature imbalances not only as problems of global public health but as a core challenge of the conservation movement, alongside habitat destruction, biodiversity loss and climate change. There is, however, little consideration of how new biosecurity concerns might alter conservation practice with unexpected and potential harmful impacts on human communities, particularly in developing nations with significant human-wildlife interfaces. Reviewing emerging policy positions from key conservation organizations, this article argues that the proposed responses to the COVID-19 pandemic hold the potential to (a) amplify existing people-park conflicts, and (b) generate new tensions by integrating global systems of viral surveillance into biodiversity conservation. I conclude that the close integration of biosecurity concerns into conservation policies requires greater acknowledgment of the unique challenges for human communities.