title: Peptide-conjugated glass slides for selective capture and purification of diagnostic cells: Applications in urine cytology.
abstract: Obtaining a clear view of the cells of interest in diagnostic cytology can be challenging when specimens are contaminated with blood or other obscuring cells. In this study, we present a powerful technique for the selective capture of diagnostic epithelial cells directly on a microscope slide, highlighting its applications in urine cytology and immunocytochemistry (ICC). Using phage-display biopanning, we identified and synthesized a series of peptides that bind with high affinity to urothelial cells but not blood cells. We developed methods for conjugating the peptides to glass slides, and we used these slides to selectively capture both normal and cancerous epithelial cells from urine contaminated with blood cells. Unlike non-selective microscope slides, the peptide-conjugated slides selectively retained the cells of interest, recovering up to 75% of urothelial cells, while up to 98% of blood cells were washed away. The slides are compatible with Papanicolaou and hematoxylin and eosin (H&E) staining for cytology preparations, as well as ICC for detecting membrane-associated and nuclear cancer markers. We successfully detected the expression of carcinoembryonic antigen and survivin, two commonly measured bladder cancer markers. In addition to bladder cancer diagnostics, this technology has broad applications for increasing the quality of sample preparations in slide-based diagnostic testing.
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Danuta B
authors.initials: DB
authors.lastname: Wronska
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Krajewska
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Natalia
authors.initials: N
authors.lastname: Lygina
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Juhua C
authors.initials: JC
authors.lastname: Morrison
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Dalia
authors.initials: D
authors.lastname: Juzumiene
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: W David
authors.initials: WD
authors.lastname: Culp
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Shrikumar A
authors.initials: SA
authors.lastname: Nair
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Martyn
authors.initials: M
authors.lastname: Darby
authors.affiliation: Affinergy, LLC, Research Triangle Park, NC.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Hofmann
mesh_terms: D001773:Blood Cells / Q000473:pathology; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis; D003581:Cytodiagnosis; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D051034:Inhibitor of Apoptosis Proteins / Q000096:biosynthesis; D009369:Neoplasms / Q000473:pathology / Q000652:urine*; D065006:Papanicolaou Test; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D000077022:Survivin; D019459:Urothelium / Q000473:pathology
chemical_list: C107489:BIRC5 protein, human; D002272:Carcinoembryonic Antigen; D051034:Inhibitor of Apoptosis Proteins; D010455:Peptides; D000077022:Survivin
keywords: bladder cancer; cell enrichment; epithelial cells; immunocytochemistry; in vitro diagnostics; peptide; urine cytology