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Susan Pang

Publications and source records attributed to Susan Pang.

5 recordsLinked to original sources
PubMed2005

A comparability study of the emerging protein array platforms with established ELISA procedures.

Recent developments in microarray technology have made it possible to perform immunoassays in a multiplexed format. This ability is highly desirable given the potential for greater throughput analysis. In spite of the obvious advantages, a number of issues arise as a result of multiplexing the reactions. In this study, we have assessed the performance characteristics that are associated with the transfer of technology from a uniplexed to a multiplexed format. Two solution phase array platforms were chosen for this study: the commercially available Luminex(100) xMap system (Austin, Texas, USA) and the UltraPlex technology devised by SmartBead Technologies Ltd. (Cambridge, UK). For this comparative study, a test for the presence of six autoantibodies in a selection of human patient serum samples was chosen as a model system. The multiplexed Luminex xMap and SmartBead UltraPlex assays were generally comparable. However, both systems generated some results that were at variance with those obtained by ELISA. The different methods used for the assignment of the cut-off levels for each of the assays within any given platform was identified as the major source of these non-concordant results. The present study demonstrates that array platforms have the potential to be used in immunodiagnostics providing that suitable cut-off levels are established.

Antibodies↗
PubMed2005

N-glycans as apical targeting signals in polarized epithelial cells.

MDCK (Madin-Darby canine kidney) cells represent a good model of polarized epithelium to investigate the signals involved in the apical targeting of proteins. As reported previously, GPI (glycosylphosphatidylinositol) anchors mediate the apical sorting of proteins in polarized epithelial cells through their interaction with lipid rafts. However, using a naturally N-glycosylated and GPI-anchored protein, we found that the GPI anchor does not influence the targeting of the protein. It is, in fact, the N-glycans that signal the protein to the apical surface. In the present review, the role of N-glycans and GPI anchors as apical signals is discussed along with the putative mechanisms involved.

Animals↗
PubMed2004

N-glycans, not the GPI anchor, mediate the apical targeting of a naturally glycosylated, GPI-anchored protein in polarised epithelial cells.

The glycosyl-phosphatidylinositol (GPI) anchor mediates the apical sorting of proteins in polarised epithelial cells through its interaction with lipid rafts. Here we investigated the signals required for the apical targeting of the naturally N-glycosylated and GPI-anchored membrane dipeptidase by selective point mutation to remove the GPI anchor addition signal or the sites for N-linked glycosylation, or both. Activity assays, immunoblotting and immunofluorescence microscopy revealed that the constructs lacking the GPI anchor were secreted from Madin-Darby canine kidney (MDCK) cells, whereas those retaining the GPI anchor were attached at the cell surface, irrespective of the glycosylation status. Wild-type membrane dipeptidase was expressed preferentially on the apical surface of both MDCK and CaCo-2 cells. By contrast, the GPI-anchored construct lacking the N-glycans was targeted preferentially to the basolateral surface of both cell types. In constructs lacking the GPI anchor, the N-glycans also targeted the protein to the apical surface. Both the apically targeted, glycosylated and the basolaterally targeted, unglycosylated GPI-anchored forms of the protein were located in detergent-insoluble lipid rafts. These data indicate that it is the N-glycans, not the association of the GPI anchor with lipid rafts, which determine apical targeting of an endogenously N-glycosylated, GPI-anchored protein in polarised epithelial cells.

Animals↗
PubMed2003

Changes of angiotensin-converting enzyme activity in the pancreas of chronic hypoxia and acute pancreatitis.

Emerging data have provided evidence for the presence of a local renin-angiotensin system (RAS) in the pancreas, which play a role in the regulation of pancreatic microcirculation, thus affecting islet hormonal secretion. The present study aimed, therefore, at elucidating the presence and changes of angiotensin-converting enzyme (ACE) using reverse transcription-polymerase chain reaction (RT-PCR) and a specific assay for ACE activity using the internally quenched fluorogenic substrate Meoc-DL-Amp-Gly-Lys(epsilon -DNP)-Gln-OH. RT-PCR clearly demonstrated the expression of ACE mRNA in the pancreas. ACE activity was markedly and significantly increased by chronic hypoxia and by acute pancreatitis when compared with that of their respective control pancreas. Addition of captopril, a specific inhibitor for ACE, completely blocked the ACE activity both in the control and experimental groups. All these data suggest that increased activity of pancreatic ACE in chronic hypoxia and acute pancreatitis could have implications for pancreatic physiology and pathophysiology.

Acute Disease↗