A pilot study to assess reproducibility of 1,305 proteins in peritoneal fluid

Principal Investigator: Alexa D. Poremba

Authors: Alexa D. Poremba, Simon T. Dillon, Allison F. Vitonis, Ana Babic, Naoko Sasamoto, Towia A. Libermann, Kathryn L. Terry
Lay Abstract

In our study, we tested the performance of a cutting edge biomarker platform by measuring 1,305 proteins in peritoneal fluid samples (fluid collected from the abdomen during surgery). For most proteins, results were similar for duplicate samples suggesting this platform provides reproducible results and can be used to study these proteins in peritoneal fluid samples. These results provide support for the use of this platform to use these peritoneal fluid samples to further study and understand endometriosis (a condition that causes pelvic pain because of the appearance of endometrial tissue on the outside of the uterus).

Scientific Abstract

Objective: Assess the reproducibility of protein levels measured in peritoneal fluid using a highly multiplexed aptamer-based platform to facilitate identification of biomarkers for endometriosis.

Materials and Methods: Proteins in peritoneal fluid samples were measured on the SOMAscan platform in the Dana-Farber/Harvard Cancer Center Cancer Proteomics Core. The plasma kit was run at 20%, 0.5% and 0.01% dilutions. 50ul of volume per sample was used to simultaneously quantify 1,305 proteins, including various inflammatory chemokines, cytokines, growth factors, immune markers, soluble receptors, and hormones. We assessed assay performance by comparing values of blinded quality control aliquots. We estimated assay variability by calculating the intra-assay and inter-assay coefficients of variation (CVs; standard deviation over the mean) in 26 duplicate samples from 13 participants.

Results and Conclusion: Of the 1,305 proteins, 62% had CV <10% and 98% had CV <20%, with only 5 proteins having CV>25%. Measurements from this platform are highly reproducible and provide support for use of this multiplex platform to evaluate protein markers in peritoneal fluid as potential diagnostic and prognostic biomarkers for endometriosis.

Clinical Implications
The results of this pilot study will allow us to advance our goal to find biomarkers for endometriosis.

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