Ultra-fast, high throughput and inexpensive detection of SARS-CoV-2 seroconversion using Ni2+ magnetic beads.

To monitor the levels of protecting antibodies raised in the population in response to infection and/or to immunization with SARS-CoV-2, we need a technique that allows high throughput and low-cost quantitative analysis of human IgG antibodies reactive against viral antigens. Here we describe an ultra-fast, high throughput and inexpensive assay to detect SARS-CoV-2 seroconversion in humans. The assay is based on Ni2+ magnetic particles coated with His tagged SARS-CoV-2 antigens. A simple and inexpensive 96 well plate magnetic extraction/homogenization process is described which allows the simultaneous analysis of 96 samples and delivers results in 7 min with high accuracy.

Magnetic Bead-Based Immunoassay Allows Rapid, Inexpensive, and Quantitative Detection of Human SARS-CoV-2 Antibodies.

Immunological methods to detect SARS-CoV-2 seroconversion in humans are important to track COVID-19 cases and the humoral response to SARS-CoV-2 infections and immunization to future vaccines. The aim of this work was to develop a simple chromogenic magnetic bead-based immunoassay which allows rapid, inexpensive, and quantitative detection of human antibodies against SARS-CoV-2 in serum, plasma, or blood. Recombinant 6xHis-tagged SARS-CoV-2 Nucleocapsid protein was mobilized on the surface of Ni2+ magnetic beads and challenged with serum or blood samples obtained from controls or COVID-19 cases. The beads were washed, incubated with anti-human IgG-HPR conjugate, and immersed into a solution containing a chromogenic HPR substrate. Bead transfer and homogenization between solutions was aided by a simple low-cost device. The method was validated by two independent laboratories, and the performance to detect SARS-CoV-2 seroconversion in humans was in the same range as obtained using the gold standard immunoassays ELISA and Luminex, though requiring only a fraction of consumables, instrumentation, time to deliver results, and volume of sample. Furthermore, the results obtained with the method described can be visually interpreted without compromising accuracy as demonstrated by validation at a point-of-care unit. The magnetic bead immunoassay throughput can be customized on demand and is readily adapted to be used with any other 6xHis tagged protein or peptide as antigen to track other diseases.

The Slr0058 Protein From Synechocystis sp. PCC 6803 Is a Novel Regulatory Protein Involved in PHB Granule Formation.

During phases of nitrogen starvation, the photosynthetic cyanobacterium Synechocystis sp. PCC 6803 produces polyhydroxybutyrate (PHB). This polymer is of high biotechnological relevance because of its potential as biodegradable plastic. Analysis of the Synechocystis genome revealed an operon (slr0058-slr0061) containing several genes, which are putatively related to the PHB metabolism. While Slr0058 show similarities with the regulatory phasin PhaF, the protein Slr0060 could serve as an intracellular PHB depolymerase. Investigation of respective knock-out mutants showed no distinct phenotype for the strain lacking Slr0060, whereas the Δslr0058 mutant displayed a growth impairment as well as a change in pigmentation. During nitrogen starvation, the Δslr0058 mutant produced in average more than twice the amount of PHB granules per cell, while the overall amount of PHB remained the same. This indicates that Slr0058 plays a role in PHB granule formation and controls it surface-to-volume ratio. GFP-tagged Slr0058 did not co-localize with PHB granules during nitrogen starvation but aggregated in distinct foci during vegetative growth. This work helps to better understand the PHB metabolism of Synechocystis sp. PCC 6803, coming closer to a sustainable, industrial production of PHB.