Affinity-free enrichment and mass spectrometry analysis of the ovarian cancer biomarker CA125 (MUC16) from patient-derived ascites,Analyst

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Affinity-free enrichment and mass spectrometry analysis of the ovarian cancer biomarker CA125 (MUC16) from patient-derived ascites
Analyst ( IF 3.6 ) Pub Date : 2020-11-03 , DOI: 10.1039/d0an01701a
Naviya Schuster-Little ₁ , Roberta Fritz-Klaus , Mark Etzel , Niharika Patankar , Saahil Javeri , Manish S Patankar , Rebecca J Whelan

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Developing a mass spectrometry-based assay for the ovarian cancer biomarker CA125 (MUC16) is a desirable goal, because it may enable detection of molecular regions that are not recognized by antibodies and are therefore analytically silent in the current immunoassay. Additionally, the ability to characterize the CA125 proteoforms expressed by individuals may offer clinical insight. Enrichment of CA125 from malignant ascites may provide a high-quality source of this important ovarian cancer biomarker, but a reliable strategy for such enrichment is currently lacking. Beginning with crude ascites isolated from three individual patients with high grade serous ovarian cancer, we enriched for MUC16 using filtration, ion exchange, and size exclusion chromatography and then performed bottom-up proteomics on the isolated proteins. This approach of enrichment and analysis reveals that the peptides detected via mass spectrometry map to the SEA domain and C-loop regions within the tandem repeat domains of CA125 and that peptide abundance correlates with clinical CA125 counts.

中文翻译：

来自患者腹水中的卵巢癌生物标志物 CA125 (MUC16) 的无亲和富集和质谱分析

为卵巢癌生物标志物 CA125 (MUC16) 开发基于质谱的分析是一个理想的目标，因为它可以检测抗体无法识别的分子区域，因此在当前的免疫分析中分析沉默。此外，表征个体表达的 CA125 蛋白型的能力可能会提供临床洞察力。从恶性腹水中富集 CA125 可能提供这种重要的卵巢癌生物标志物的高质量来源，但目前缺乏这种富集的可靠策略。从从三名高级别浆液性卵巢癌患者中分离的粗腹水开始，我们使用过滤、离子交换和尺寸排阻色谱法富集 MUC16，然后对分离的蛋白质进行自下而上的蛋白质组学。通过质谱法映射到 CA125 串联重复结构域内的 SEA 结构域和 C 环区域，并且肽丰度与临床 CA125 计数相关。

更新日期：2020-11-03

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