A new electrochemical assay for the detection of secreted alkaline phosphatase (SEAP) from transfectant HeLa cells is proposed using a microarray device and scanning electrochemical microscopy (SECM). The assay consists of two steps: the first is the incubation of a transfected cell in a microarray culture device covered with a substrate modified with anti-SEAP under physiological conditions without any additives. The array device consists of a 4 × 4 array of microwells having a size of 100 µm × 100 µm (diameter × depth). The second step is SECM measurement of secreted SEAP at the antibody-immobilized substrate. This assay ensures accuracy and intactness because the undesired influence of endogeneous ALP is eliminated and the transfected cells are incubated in a culture device under suitable conditions. We successfully detected the expression of SEAP from intact cells at the single-cell level using this assay. The system is useful as a cell-based gene-expression assay. Biotechnol. Bioeng.
Single-cell analysis has become a powerful method to construct a new type of whole cell sensor integrating with individual cellular responses all together. However, sensitivity of electrochemical detection system in the present stage is insufficient to discuss in detail about the results obtained...
A dual reporter cell assay (DRCA) that allows real-time detection of herpes simplex virus (HSV) infection was developed. This was achieved by stable transfection of cells with an expression cassette that contains the dual reporter genes, the secreted alkaline phosphatase (SEAP) and the enhanced g...
A cell-based bioassay capable of detecting neutralizing antibodies (NAb) specific to a therapeutic anti-IL-13 monoclonal antibody was developed, validated and used to analyze normal human and asthma serum samples. At the time of this study, a neutralizing assay was unavailable for anti-IL-13 anti...
Activation of intestinal epithelial cell (IEC) nuclear factor IoB (NFIoB) and the consequent chemokine upregulation are crucial events in inflammatory bowel disease (IBD) pathogenesis. Not much is known about the consequences of NFIoB inhibition in terms of chemokine expression in intestinal cell...
Adenovirus CELO is a promising vector for gene delivery to eukaryotic cells including mammalian cells. It allows one to use recombinant CELO adenoviruses against animal pathogens. A high expression level of transgene in the viral vector is necessary to reach the humoral and T-cell immune answer. ...
submitted by wentupthehill 2 years and 2 months ago
Novel mixed-lineage kinase protein zipper sterile-[alpha]-motif kinase (ZAK) was first cloned by our laboratory. Lung cancer is the leading cause of cancer-related death in the world, including in Taiwan. Here, we wanted to investigate whether ZAK plays a potential role in lung cancer development...
Fast and efficient production of recombinant proteins for structural and functional studies is a crucial issue for research and for industry. To this end, we have developed an efficient system to generate in less than 2 months, starting from the cDNA, pools of CHO cells stably expressing high-lev...
submitted by niaraindacreni 1 year and 9 months ago
Improvements are needed for the early detection of breast cancer, as current imaging methods lack sensitivity to detect small tumors and assess their disease phenotype. ...
submitted by corlahomilenck 1 year and 9 months ago
MRNA from single cells was quantified using real-time RT-PCR after recording the address and reporter protein activity with chemiluminescence, fluorescence, and electrochemical techniques, using luciferase, green fluorescent protein, and secreted alkaline phosphatase. mRNA copy number ranging fro...
Productivity and sialylation are two important factors for the production of recombinant glycoproteins in mammalian cell culture. In our previous study, we found that silkworm hemolymph (SH) increased the sialylation of recombinant secreted human placental alkaline phosphatase (SEAP) in the insec...
