Design and Development of Lateral Flow Test for Early Screening of Pancreatic Cancer

Abstract

Pancreatic cancer currently showing one of the lowest survival rates in the UK (~5%). Despite the huge efforts in improving the diagnosis using existing screening methods, an early stage detection remains a challenge. Developing a rapid, point-of-care lateral flow test (LFT) for patients is an unmet clinical goal that can accelerate the screening in clinical settings. The focus of my project is to design & develop a LFT based on carbohydrate/cancer antigen (CA 19-9) levels in the blood. My aim is to investigate and increase the sensitivity of the LFT by indicating high-low levels of CA 19-9 for regular screening and monitoring. This project involved 4 distinct stages: Optimizing the appropriate golden nanoparticles (Au NP) for their functionalization with different concentrations of PEG and MSA to get the best immunoassay. Different characterization methods including UV-Vis and DLS will be conducted to MSA and PEG Au NPs to observe the stability of Au NPs at different concentrations. Consequently, freeze-drying procedure will be conducted to ensure that PEG and MSA AuNPs sensitivity and reactivity is retained when in dried state and rehydration/rewetting. All MSA AuNP aggregated after resuspension. While 50 uM, and 100 uM PEG retained stability after resuspension. 50 uM PEG was selected for the functionalization of 22 nm AuNP (AuNP1) for further Anti-CA 19-9 monoclonal antibody covalent conjugation. Characterisation methods including UV-VIS, DLS, gel electrophoresis (agarose and ) indicated successful EDC/NHS conjugation. To assess the binding efficiency of Ab-AuNP on membrane strip; Supernatant was collected from cholangiocarcinoma cell lines and pancreatic cell lines at 80% confluency. The expression of CA 19-9 was evident in PDX-185 only. No test line were evident; during suspension of SFM PD-185 supernatant and Ab-AuNP1 on immobalised mAb. Hence, the Fabrication of the LFT Lastly was not achieved as Ab concentration optmisation during conjugation process is required to achieve the optimum sensitivity and the limit of detection for the LFT.