title: Interrogating Bronchoalveolar Lavage Samples via Exclusion-Based Analyte Extraction.
abstract: Although average survival rates for lung cancer have improved, earlier and better diagnosis remains a priority. One promising approach to assisting earlier and safer diagnosis of lung lesions is bronchoalveolar lavage (BAL), which provides a sample of lung tissue as well as proteins and immune cells from the vicinity of the lesion, yet diagnostic sensitivity remains a challenge. Reproducible isolation of lung epithelia and multianalyte extraction have the potential to improve diagnostic sensitivity and provide new information for developing personalized therapeutic approaches. We present the use of a recently developed exclusion-based, solid-phase-extraction technique called SLIDE (Sliding Lid for Immobilized Droplet Extraction) to facilitate analysis of BAL samples. We developed a SLIDE protocol for lung epithelial cell extraction and biomarker staining of patient BALs, testing both EpCAM and Trop2 as capture antigens. We characterized captured cells using TTF1 and p40 as immunostaining biomarkers of adenocarcinoma and squamous cell carcinoma, respectively. We achieved up to 90% (EpCAM) and 84% (Trop2) extraction efficiency of representative tumor cell lines. We then used the platform to process two patient BAL samples in parallel within the same sample plate to demonstrate feasibility and observed that Trop2-based extraction potentially extracts more target cells than EpCAM-based extraction.
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: Jacob J
authors.initials: JJ
authors.lastname: Tokar
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: Jay W
authors.initials: JW
authors.lastname: Warrick
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Guckenberger
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: Jamie M
authors.initials: JM
authors.lastname: Sperger
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: Joshua M
authors.initials: JM
authors.lastname: Lang
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: J Scott
authors.initials: JS
authors.lastname: Ferguson
authors.affiliation: 1 University of Wisconsin Madison, Madison, WI, USA.|2 UW Carbone Cancer Center, Madison, WI, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology*; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry / Q000295:instrumentation / Q000379:methods*; D008175:Lung Neoplasms / Q000175:diagnosis*; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods*
chemical_list: D014408:Biomarkers, Tumor
keywords: bronchoalveolar; clinical automation; lab on a chip; lavage; lung cancer; microtechnology; solid-phase extraction