Unique multiarray immunosensor for the accurate quantification of the fertility window of women in saliva

Project Information

Umay4women

Grant agreement ID: 101024231

DOI

10.3030/101024231

Project terminated on 31 March 2023

EC signature date 6 April 2021

Start date 1 September 2021

End date 31 August 2023

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 178 320,00

EU contribution

€ 178 320,00

Coordinated by

UNIVERSITEIT ANTWERPEN
Belgium

Periodic Reporting for period 1 - Umay4women (Unique multiarray immunosensor for the accurate quantification of the fertility window of women in saliva)

Reporting period: 2021-09-01 to 2023-08-31

Summary of the context and overall objectives of the project

According to WHO, ca. 8 – 10 % of couples (50 million people) are facing fertility problems. The quantification and monitoring of specific female hormones are crucial for the early identification of infertility and the tracing of diseases associated with hormonal disbalance (e.g. ovarian cancer). One of the main reasons couples have difficulty conceiving is their inability to accurately predict the female’s ovulation period. In comparison with costly and complex conventional methods and commercially available tests that only measure one or two of the four key hormones, Umay proposed a unique and reliable quantification of all hormones involved in the ovulatory cycle to accurately determine the ‘fertility window’ of women by using non-invasive saliva samples. The novelty of this proposal relied on the combination of nanomaterials, photosensitisers, paper-based microfluidics, and immunoassay disciplines to develop a biosensor that can detect and quantify the key hormones associated with fertility in women, overcoming the drawbacks of current techniques and sampling methods. Although initially focused on fertility monitoring in women, the underlying technologies have the potential to be further extended after this fellowship for a wider range of applications and final users (e.g. monitoring of fertility in the animal industry or tracing the evolution of patients after ovarian cancer treatment) to develop reliable, low-cost, multiarray platforms for healthcare applications. From the clinical perspective, Umay will facilitate the direct and rapid quantification of the key fertility hormones which would lead to faster and private decision-making processes concerning the fertility status of each woman.

Umay’s main goal was the simultaneous quantification of four key hormones involved in the ovulatory cycle (in saliva) allowing for the identification of the fertility window specific for each woman in an accurate and non-invasive way. The specific research objectives pursued to accomplish the main goal were:
a) Fabrication and development of a novel POC platform based on the combination of paper microfluidic and electrochemical sensors using low-cost and scalable screen-printing technology; work package (WP) 2.
b) Develop an innovative sensing strategy by functionalisation of the biosensor with antibodies and innovative photosensitisers-conjugated elements that can selectively and sensitively detect each hormone; WP 3.
c) Integration of the multiarray biosensor in the microfluidic paper-based platform enabling the detection of several hormones in a single assay by the embodiment in a 3D printed device; WPs 3-4.
d) Non-invasive quantification and validation of the selected hormones using saliva samples; WP 4.

The data generated during this action will help in the understanding and future development of the ideal Umay multiarray sensor.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

Several publications, collaborations, and experimental activities have been performed.

1. Three peer-reviewed articles and one review were published in high-impact journals:
a) Electrochemical identification of hazardous phenols and their complex mixtures in real samples using unmodified screen-printed electrodes. H Barich, R Cánovas, K De Wael. Journal of Electroanalytical Chemistry 904, 115878.
b) Novel electrochemiluminescent assay for the aptamer-based detection of testosterone. R Cánovas, E Daems, R Campos, S Schellinck, A Madder, JC Martins, F Sobott, K De Wael. Talanta 239, 123121.
c) Development of a combi-electrosensor for the detection of phenol by combining photoelectrochemistry and square wave voltammetry. L Neven, H Barich, N Sleegers, R Cánovas, G Debruyne, K De Wael. Analytica Chimica Acta 1206, 339732.
d) Are aptamer-based biosensing approaches a good choice for female fertility monitoring? A comprehensive review. R Cánovas, E Daems, AR Langley, K De Wael. Biosensors and Bioelectronics, 114881.
These publications were directly related to the content of the action since all of them are focused on the photoelectrochemical detection of phenolic compounds (used as a mediator of the singlet-oxygen reaction needed to boost the response) and/or on the detection of the hormones of interest (progesterone, estradiol, luteinizing hormone or follicle stimulating hormone) and other small steroids such as testosterone. Part of the research was also disseminated during the attendance to the conferences: CSAC2021 online presentation and BES2022 poster presentation.

2. (Inter)national collaborations with: Prof. Sofie Cambré (Physics department, UAntwerpen) specialised in carbon nanotubes, Prof. Sarah Lebeer (Bioengineering department, UAntwerpen) centred on women’s health, MD. Timon Vandamme (oncologist from UZA), Dr. Eva Colas (translational research group in Vall Hebron Research Institute, Barcelona) focused on investigating endometriosis, and MD. Lidia Melero, gynaecologist-oncologist and surgeon from Virgen del Rocio University Hospital (Sevilla).

3. The experimental part can be summarised into the investigation and development of:
• Home-made screen-printed electrodes (SPEs) using our screen-printing facilities and their characterisation via cyclic, square wave, and linear sweep voltammetric techniques (WP2).
• After the fabrication of the SPEs, the next steps were (WP3):
a) the use of different nanomaterials (i.e. gold nanoparticles, carbon nanotubes, and magnetic beads) as well as their optical and electrochemical characterisation.
b) The functionalisation of the working electrodes with the biorecognition elements that will enable the detection of the hormones via sandwich assay and photoelectrochemistry. This sandwich assay contains a primary antibody, the hormone of interest, and the secondary biorecognition element labelled with a photosensitiser needed for triggering the response.
c) Finally, the analytical performance of the sandwich assay was tested by performing several experiments to optimise the design of the assay depending on the concentration of the mediator (hydroquinone), time of incubation, illumination, applied potential, nanomaterials, etc.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

All current methods of fertility monitoring are characteristically invasive, which can cause pain and discomfort to the patient. Therefore, a rapid, at-home fertility monitoring tool would be ideal to alleviate the apprehensiveness associated with routine screenings and give women the privacy desired when trying to conceive.

Moreover, although the project was initially focused on fertility monitoring in women, the underlying technologies have the potential (and socio-economic impact) to be further extended for a wider range of applications and final users (e.g. monitoring of fertility in the animal industry: dog breeders, cow farms, horses, etc. or tracing the evolution of patients after ovarian cancer treatment, a “daughter” line collaboration that is being initiated thanks to Umay) to develop reliable, low-cost, multiarray platforms for healthcare applications.

The innovative aspect of the research programme is also related to its impact on society as the ideal Umay sensor could help the health sector to detect possible infertility earlier, hence improving the quality of life of couples interested in conceiving. From the clinical perspective, Umay would facilitate the direct and rapid quantification of the key fertility hormones which will lead to faster and private decision-making processes concerning the fertility status of each woman.

Umay sensor concept