Navina Lotay

Lysophosphatidic acid (LPA) is a cell-signaling lipid that has been proposed as an early-stage biomarker for ovarian cancer (OC). Diagnosing OC in Stage I is critical to improving patient outcomes, increasing the survival rate from 30% (when diagnosed in late stages of the disease) to over 90%. This significant improvement is due to the success of early interventions; however, current diagnostic methods are not as effective at early-stage detection, with only 15% of cases diagnosed in Stage I and over 70% diagnosed in Stage III or IV. There is a strong need for LPA detection that is sensitive, specific, rapid, low-cost, and automated to truly validate its effectiveness as a diagnostic characteristic for OC. We report the preliminary development and characterization of one such biosensor, which makes use of the advantages of magnetic particles and chemiluminescence for quick, sensitive detection of LPA. The sensor has proven to be viable, with a positive response to LPA concentration, a measurement time of 5 s after incubation, and an LOD of 3.5 nM.

Carbon dots (CDs) are versatile, easy to fabricate, chemically stable, and ecofriendly materials. This work presents the impact of different post-treatments, such as centrifugation and filtration, on the photoluminescence (PL) and particle-size distribution of electrochemically synthesized CDs. Because organic envelope surfactants have been reported to influence PL, bare CDs were compared with CDs modified with amphiphilic and nonamphiphilic surfactants. The CD colloidal solution exhibited stabilization in the case of bare CDs and CDs modified with amphiphilic sodium dodecyl sulphate or non-amphiphilic surfactant sodium citrate, as indicated by the zeta potential. The experimental results also revealed a minor influence of the organic envelope on the absorbance or PL spectrum, whereas filtration of the colloid solution had a significant impact, revealing an effect not previously observed. The change in the absorbance spectrum was attributed to sp³ hybridization due to increased disorder and surface states during the filtration process. Absorbance deconvolution also disclosed two energy substates not mentioned elsewhere. The PL spectra of the filtered CD solutions revealed that the emission peaks shifted according to the incident wavelength, and the centrifuged CDs synthesized with the sodium citrate surfactant lacked PL due to the absence of sp³ states. This study emphasizes the importance of CD post-treatment in CD production rather than the use of surfactants.

Gelsolin is an actin-binding protein that is competitively bound by lysophosphatidic acid (LPA), a possible biomarker for the early detection of ovarian cancer. Our group has previously shown the usage of gelsolin 1-3 (a fragment of gelsolin composed of the first three of its six subdomains) along with actin to detect the presence of lysophosphatidic acid. This histidine-tagged gelsolin 1-3 fragment is synthesized in our lab using bl21 Rosetta cells but required further characterization. This study aims to provide sufficient characterization of gelsolin 1-3 synthesized in our lab via both sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry using a Fourier-transform ion cyclotron resonance mass spectrometer (FTICR-MS). Both techniques show the presence of protein at the expected mass of approximately 41 kDa. Additionally, the dissociation of a smaller fragment of the complex when introduced into the gas phase which was equivalent to the approximate mass of one subdomain of gelsolin 1-3 further supports this conclusion.

The overall 5-year survival rate of ovarian cancer (OC) is generally low as the disease is often diagnosed at an advanced stage of progression. To save lives, OC must be identified in its early stages when treatment is most effective. Early-stage OC causes the upregulation of lysophosphatidic acid (LPA), making the molecule a promising biomarker for early-stage detection. An LPA assay can additionally stage the disease since LPA levels increase with OC progression. This work presents two methods that demonstrate the prospective application for detecting LPA: the electromagnetic piezoelectric acoustic sensor (EMPAS) and a chemiluminescence-based iron oxide nanoparticle (IONP) approach. Both methods incorporate the protein complex gelsolin–actin, which enables testing for detection of the biomarker as the binding of LPA to the complex results in the separation of gelsolin from actin. The EMPAS was characterized with contact angle goniometry and atomic force microscopy, while gelsolin–actin-functionalized IONPs were characterized with transmission electron microscopy and Fourier transform infrared spectroscopy. In addition to characterization, LPA detection was demonstrated as a proof-of-concept in Milli-Q water, buffer, or human serum, highlighting various LPA assays that can be developed for the early-stage detection of OC.

Gynecological medicine is a crucial field that is limited by standard methods of screening and detection. Identification of biomarkers and sensor development for women's cancers and illnesses is a vast field that aims to improve the health of women.