FastEV & Lectin
FastEV unlocks the cancer biomarker cargo of serum
Pooled serum samples from cancer patients or controls were fractionated with an array of FastEV conditions to reveal distinct enrichments of cancer-linked targets by a lectin -based platform at University of Turku. Both FastEV isolates and supernatants were subjected to analysis.
Methods: Analysis of FastEV fractions with lectin platform
Starting material in FastEV was 120 µl of pooled serum/condition and isolations were done on 96-well plates. Total protein was measured in FastEV isolates and supernatants with Nanodrop (Thermo Fischer Scientific) and equal protein quantities applied to lectin platform.
Figure: Distribution of cancer-linked markers in FastEV and supernatant fractions. Signals from the lectin-based platform varied according to FastEV conditions, fractions (EV or supernatant) and cancer /condition type. Specific EV fractions enriched e.g. AOL and UEA, while other targets were more variably enriched in the EVs or supernatants, e.g. CA19-9. Bubble size indicates signal strength. Data is from two technical replicates per assay and sample. EV, FastEV isolate; SP, supernatant; BlCa, Bladder Cancer; BPH, Benign Prostate Hyperplasia; Endo, Endometrioid Carcinoma; OvCa, Ovarian carcinoma.
Identification of optimal FastEV conditions for biomarker discovery in bladder and ovarian cancer
FastEV was used to fractionate serum to find conditions bringing out the greatest differences between cancer and control samples in the lectin platform. The screen successfully identified specific FastEV conditions with significant potential in biomarker discovery for ovarian and bladder cancer.
Figure: Specific FastEV conditions show high potential for separating cancer from control in the lectin platform. Fold change (FC) of specific cancer-linked signals between cancer vs control were high from two FastEV conditions (one EV and one SP) in bladder cancer vs benign prostate hyperplasia. Even higher fold changes were observed from the top conditions (EV and SP) of ovarian cancer vs endometriosis assay. Data is from serum pools, where N=10 of individual samples of pool for all groups. Two technical replicates were analyzed per assay and sample. EV, FastEV isolate; SP, supernatant; BlCa, Bladder Cancer; OvCa, Ovarian carcinoma.