Biology subjects

Adams, C. T.

Publications and source records attributed to Adams, C. T..

3 recordsLinked to original sources

bioRxiv2025

Single-molecule validation and optimized protocols for the use of secondary nanobodies in multiplexed immunoassays

Recently developed secondary nanobodies or single-domain antibodies present a powerful tool for immunodetection. Unlike traditional antibodies, their monovalence enables pre-association with primary antibodies prior to sample staining, presenting a straightforward affinity-based antibody labeling solution. This not only simplifies and streamlines immunoassays, it also supports multiplexed techniques where conflicts in the species of the desired primary antibodies preclude standard indirect immunostaining. Despite these advantages, the use of secondary nanobodies remains sparse, due perhaps to a lack of evaluation on their suitability for assays requiring quantification and an assessment of optimal protocols for their use. Here, we present a set of experiments spanning single-molecule detection to cell imaging that can be used to validate secondary nanobody binding, specificity, and their propensity for mis-targeted binding in multiplex assays. Using these tools, we analyzed the binding properties of commercially available secondary nanobodies and outline optimized protocols for their robust use.

biochemistry↗

bioRxiv2024

Analyzing Biodiversity in Protected vs. Unprotected Tide Pools

This study assessed the efficacy of marine protected areas (MPAs) by comparing tide pool biodiversity in Half Moon Bay, California between a protected site (Fitzgerald Marine Reserve) and an unprotected site (Mavericks Beach). Samples were collected through random quadrat surveys. ImageJ and RStudio were used to analyze data through three methods utilizing the Simpsons Diversity Index along with three metrics. The unprotected Mavericks Beach displayed greater biodiversity through two metrics while the third did not find significant difference in biodiversity between sites. 30 unique species were present at each site with 43 species between both. These mixed findings prevent us from drawing definitive conclusions on the effectiveness of Half Moon Bays tide pool MPAs but suggest the MPA is less biodiverse. Potential explanations include the Intermediate Disturbance Hypothesis, lack of sample size, or that the MPA is not effective in maintaining biodiversity.

ecology↗

bioRxiv2024

Testing a Method for Quantifying Microplastic Content in Mytilus californianus

With the proliferation of microplastics within the environment, there has been an increased need to quantify them within organic tissues of aquatic species. Several studies have examined the potential role of mussels in filtering these contaminants, prompting interest in characterizing uptake and retention of microplastics in these species. There are a variety of methods to isolate and quantify microplastics, including density separation, enzymatic digestion, and filtration. The applicability of a combined digestion and ultrasonic bath methodology in isolating microplastics from the California mussel (Mytilus californianus Conrad, 1837) was investigated. Microplastics were added to 20-gallon tanks at 0 g (control) 8 mg. Mussel tissues from each tank were digested using potassium hydroxide solutions. These digestions were subjected to vacuum filtration and a sonication bath, then treated with a mixture of sodium hypochlorite and nitric acid. It was found that this method successfully removed organic tissue from the mussels while leaving dyed microplastics intact. The latter were quantified using fluorescent microscopy. This proof-of-principle experiment indicates that this technique can be applied to further studies on environmental impact and retention of microplastics in mussels and may also have utility for other shellfish.

zoology↗