Open AccessAn innovative cell selection approach in developing human cells overexpressing aspartyl/asparaginyl β-hydroxylase
Author(s) -
Hadi Bakhtiari,
Abbas Ali Palizban,
Hossein Khanahmad,
Mohammad Reza Mofid
Publication year - 2020
Publication title -
research in pharmaceutical sciences
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.685
H-Index - 29
eISSN - 1735-9414
pISSN - 1735-5362
DOI - 10.4103/1735-5362.288436
Subject(s) - transfection , recombinant dna , hek 293 cells , cell culture , microbiology and biotechnology , chemistry , flow cytometry , hygromycin b , immunocytochemistry , cell , biology , gene , biochemistry , plasmid , genetics , endocrinology
Aspartyl/asparaginyl β-hydroxylase (ASPH) is abundantly expressed in malignant neoplastic cells. The establishment of a human cell line overexpressing ASPH could provide the native-like recombinant protein needed for developing theranostic probes. In the process of transfection, the obtained cells normally contain a range of cells expressing the different levels of the target of interest. In this paper, we report on our simple innovative approach in the selection of best-transfected cells with the highest expression of ASPH using subclone selection, quantitative real-time polymerase chain reaction, and gradual increment of hygromycin concentration.