In vitro selection of CD70 binding aptamer and its application in a biosensor design for sensitive detection of SKOV-3 ovarian cells

Abstract

Single-stranded DNA aptamers recognizing CD70 molecule overexpressed in some tumor cell lines was isolated by means of systematic evolution of ligands by exponential enrichment (SELEX). Both purified CD70 protein and CD70-expressing cells were used to isolate target-specific aptamer. After certain rounds of protein-based SELEX and cell-SELEX (ten and seven rounds, respectively), Aptamer 928 (hereafter Apt928) with high affinity and specificity for CD70 was obtained. The specific binding of the aptamer to its target could block interaction of CD70 with CD27 (known as CD70 receptor). Dissociation constant of Apt928 was calculated to be 66 nmol L−1. Moreover, ATTO 674N-labeled Apt928 was applied as a fluorescent aptasensor for rapid and sensitive detection of SKOV-3 cells as CD70-positive cells. The detection limit of this aptasensor was 14 cells mL−1.