A competitive lateral flow assay for the detection of tenofovir.

Proper management of an HIV infection requires that a patient be at least 80-95% adherent to a prescribed drug regimen to avoid poor health outcomes and the development of drug-resistant HIV strains. Clinicians generally monitor adherence habits indirectly through patient self-reporting, pill counting, and electronic drug monitoring. While direct measurement of patient samples like urine for monitoring drug levels is possible, it requires specialized equipment and training that is not readily available in resource-limited settings where the need is greatest. In this work we report the development of an antibody that binds to tenofovir (TFV), a key small molecule drug for both the treatment and prevention of HIV, and a competitive lateral flow assay that uses that antibody to monitor urine samples for the presence of the drug. TFV was conjugated to an immunogenic protein and injected into rabbits to raise polyclonal antibodies sensitive to the drug. The antibodies were verified for TFV-sensitivity by immunoprecipitation and HPLC. A gold nanoparticle-based competitive assay was developed to detect the presence of TFV in urine samples with a sensitivity of 1 µg mL-1. This TFV assay could be deployed as a point-of-care device for adherence monitoring in resource-limited settings as a low-cost, accurate, and speedy alternative to current methods to better inform changes in treatment.

An Engineered Human Fc-Mannose-Binding-Lectin Captures Circulating Tumor Cells.

Tumor cells circulating throughout the body have shown great potential for providing new diagnostic or therapeutic strategies for treating cancer patients. However, isolating circulating tumor cells (CTCs) is still challenging due to the lack of broad spectrum reagents that bind specifically to these cells. This study shows that an engineered human blood opsonin that mimics the innate immune mechanism for opsonizing complex mannan carbohydrates, Fc-mannose binding lectin (FcMBL), exhibits a broad spectrum of CTC binding activity. Using FcMBL-coated magnetic beads, this study is able to specifically capture and isolate a broad range of tumor cells spiked into buffer or blood. FcMBL is bound preferentially to human and mouse breast cancer cells relative to normal breast epithelium, and this study demonstrates the capture of seven different types of cancer cells with greater than 90% capture efficiency, whereas two of these same cancer cells bound poorly to anti epithelial cell adhesion molecule antibodies. It is also confirmed that FcMBL-coated magnetic beads can be used to capture CTCs from the blood of mice bearing metastatic tumors. The FcMBL capture technology may therefore provide a new tool for harvesting a broad range of CTCs with high efficiency as it targets tumor cell specific surface markers that are expressed across diverse cell types and retained throughout the metastatic process.

Optical clearing agent perfusion enhancement via combination of microneedle poration, heating and pneumatic pressure.

BACKGROUND AND OBJECTIVE: Optical clearing agents (OCAs) have shown promise for increasing the penetration depth of biomedical lasers by temporarily decreasing optical scattering within the skin. However, their translation to the clinic has been constrained by lack of practical means for effectively perfusing OCA within target tissues in vivo. The objective of this study was to address this limitation through combination of a variety of techniques to enhance OCA perfusion, including heating of OCA, microneedling and/or application of pneumatic pressure over the skin surface being treated (vacuum and/or positive pressure). While some of these techniques have been explored by others independently, the current study represents the first to explore their use together. STUDY DESIGN/MATERIALS AND METHODS: Propylene glycol (PG) OCA, either at room-temperature or heated to 45°C, was topically applied to hydrated, body temperature ex vivo porcine skin, in conjunction with various combinations of microneedling pre-treatment (0.2 mm length microneedles, performed prior to OCA application), vacuum pre-treatment (17-50 kPa, performed prior to OCA application), and positive pressure post-treatment (35-172 kPa, performed after OCA application). The effectiveness of OCA perfusion was characterized via measurements of transmittance, reduced scattering coefficient, and penetration depth at a number of medically-relevant laser wavelengths across the visible spectrum. RESULTS: Topical application of room-temperature (RT) PG led to an increase in transmittance across the visible spectrum of up to 21% relative to untreated skin. However, only modest increases were observed with addition of various combinations of microneedling pre-treatment, vacuum pre-treatment, and positive pressure post-treatment. Conversely, when heated PG was used in conjunction with these techniques, we observed significant increases in transmittance. Using an optimal PG perfusion enhancement protocol consisting of 45°C heated PG + microneedle pre-treatment + 35 kPa vacuum pre-treatment + 103 kPa positive pressure post-treatment, we observed up to 68% increase in transmittance relative to untreated skin, and up to 46% increase relative to topical RT PG application alone. Using the optimal PG perfusion enhancement protocol, we also observed up to 30% decrease in reduced scattering coefficient relative to untreated skin, and up to 20% decrease relative to topical RT PG alone. Finally, using the optimal protocol, we observed up to 25% increase in penetration depth relative to untreated skin, and up to 23% increase relative to topical RT PG alone. CONCLUSIONS: The combination of heated PG, microneedling pre-treatment, vacuum pre-treatment, and positive pressure-post treatment were observed to significantly enhance the perfusion of topically applied PG. Although further studies are required to evaluate the efficacy of combined perfusion enhancement techniques in vivo, the current results suggest promise for facilitating the translation of OCAs to the clinic.