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Global Health Technologies

Undergraduate Lightning Talks

Optimization of an Anti-TOP2A Antibody-gold Nanoparticle Conjugation System for Point-of-Care Cervical Cancer Screening

Saturday, October 26, 2024
1:50 PM – 1:55 PM EST
Location: Room 342

Introduction:  Cervical cancer is one of the most common cancers in women globally; however, it is one of the highly treatable diseases if detected early. In lower-income communities there is limited access to preventive and screening measures hence there is a need for an early stage cervical cancer rapid diagnostic test. In cervical cancer patients, topoisomerase II alpha (TOP2A) is highly expressed, making the biomarker a good target for a rapid pre-cervical cancer rapid diagnosis. TOP2A is an enzyme involved in DNA replication, affecting the structure of DNA through interaction with the double helix. This work focuses on optimizing a conjugation system involving anti-TOP2A antibodies and gold nanoparticles (AuNPs). These conjugates will serve as labeled molecules for the TOP2A lateral flow immunoassay (LFIA) device.

Materials and

Methods:  Mouse Anti TOP2A antibodies were conjugated to 40 nm AuNPs by passive adsorption. Once the antibodies were bound to the AuNPs, a blocking agent was used to omit the chance of unspecified binding. Multiple dAb-to-AuNP molar ratios were tested using commercial quality control dipsticks and dynamic light scattering (DLS) to determine the conjugation with the highest intensity and stability. The average hydrodynamic diameter and polydispersity index (PdI) were among parameters investigated in the DLS studies. Pixel intensity was quantified using the open-source ImageJ software.

Results, Conclusions, and Discussions: The Ab: AuNP ratio of 100:1 not only produced the strongest signal on the commercial dipstick, but this ratio had the highest grayscale intensity and maintained the strong signal throughout a seven day period (33.83 ± 0.89 - Figure 1, 2A, and 2C). The hydrodynamic diameter and PdI values were also the most stable during this period (54.95 ± 0.38 and 0.18, respectively - Figure 2B). Hence, the 100:1 Ab: AuNP ratio was chosen as the optimal ratio for the LFIA device. Future experiments for the assay include optimizing the test, control lines of the LFIA, and accessing the overall performance with cultured cervical cancer cells and clinical cervical swabs. Additionally, TOP2A will be combined with other biomarkers to create a sensitive, specific, and rapid diagnostic for timely pre-cervical cancer detection and treatment in low-to-middle income communities.

Acknowledgements (Optional):