期刊
CLINICAL & TRANSLATIONAL ONCOLOGY
卷 19, 期 8, 页码 976-988出版社
SPRINGER-VERLAG ITALIA SRL
DOI: 10.1007/s12094-017-1629-y
关键词
DNA aptamers; HER2; Immunoassays; Immunohistochemistry
类别
资金
- Department of Biotechnology, New Delhi, Govt. of India [BT/CP/07/NE/TBP/2010]
- IIT Guwahati
- Ministry of Human Resource Development (MHRD), Government of India
Among four selected candidates, ECD_Apt1 (having minimum a dagger G = -3.24) showed the highest binding affinity (K (d) = 6.33 +/- 0.86 nM) to Her2-ECD protein. The aptamer-protein sandwich assay showed a linear rise in chemiluminescence (at 490 nm wavelength) in the dynamic range of 100-700 nM ECD_Apt1 with a detection limit of 12.5 +/- 2.5 ng/mL. Biotinylated ECD_Apt1 showed stronger cytoplasmic staining in Her2-positive breast cancer cell lines (SKBR3) compared to Her2-negative cells (MDA MB 231, MCF7). In paraffin-embedded breast cancer tissue sections, it showed specific and selective localization in the cytoplasmic niche of malignant duct cancer cells without any cross-reactivity to fibroblasts, inflammatory cells and adipocytes. Binding assays, cytochemical and histochemical studies support ECD_Apt1 as a potential theranostic agent for Her2-positive carcinomas. ECD_Apt1 could be an effective low-cost alternative to conventional anti-Her2 antibody in solid phase immunoassays for cancer diagnosis and related applications.
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