Development of carbon nanoparticles-based soluble solid-phase immune sensor for the quantitative diagnosis of inflammation.

Quantitative immunodiagnosis is one of the most commonly used methods for in vitro diagnostics. Various bioanalytical methods have been developed to quantitatively diagnose immune analytes; however, they require blood dilution pretreatment, reaction mixing, complicated experimental steps, and can cause diagnostic errors due to the hook effect. To address this issue, we introduced a simple immunoassay based on carbon nanoparticles (CNPs). The assay was designed to have high flexibility for use in various in vitro diagnostic devices by constructing a soluble solid-phase immune sensor with high solubility using antibody-conjugated CNPs and polymer materials. Excellent performance was achieved using a free-antibody system with dual calibration. To verify the performance of this method with high reliability, canine C-reactive protein was selected as the immune analyte. Interestingly, our method efficiently mitigated the hook effect with outstanding performance in a one-step reaction without blood dilution or reaction mixing. The detection range of the target can be effectively controlled using free antibodies. Therefore, our CNP-based immunodiagnosis method may advance the commercialization of point-of-care immune biosensors.

A MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) for total N-glycan analysis.

OBJECTIVES: To develop a sensitive and quantitative method for monitoring the abnormal glycosylation of clinical and biopharmaceutical products. RESULTS: MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) was proposed for sensitive and quantitative analysis of total N-glycans. The derivatization reactions (i.e., amidation of sialic acid and incorporation of a positive charge moiety into the reducing end) dramatically increased the linearity (R(2) > 0.99) and sensitivity (limit of detection is 0.5 pmol/glycoprotein) relative to underivatized glycans. In addition, the analytical strategy was chromatographic purification-free and non-laborious process accessible to the high-throughput analyses. We used MALDI-QTaG to analyze the N-glycans of α-fetoprotein (AFP) purified from normal cord blood and HCC cell line (Huh7 cells). The total percentages of core-fucosylated AFP N-glycans from Huh7 cells and normal cord blood were 98 and 18%, respectively. CONCLUSIONS: This MALDI-MS-based glycomics technology has wide applications in many clinical and bioengineering fields requiring sensitive, quantitative and fast N-glycosylation validation.

Comparative analysis of gene expression under cold acclimation, deacclimation and reacclimation in Arabidopsis.

Cold acclimated plants show an elevated tolerance against subsequent cold stress. Such adaptation requires alterations in gene expression as well as physiological changes. We were interested in gene expression changes at the transcriptional level during adaptation processes. The patterns of transcriptional changes associated with cold acclimation, deacclimation and reacclimation in Arabidopsis leaves were characterized using the Coldstresschip. Gene expression profiles were further analyzed by 'coexpressed gene sets' using gene set enrichment analysis (GSEA). Genes involved in signal transduction through calcium, and cascades of kinases and transcription factor genes, were distinctively induced in the early response of cold acclimation. On the other hand, genes involved in antioxidation, cell wall biogenesis and sterol synthesis were upregulated in the late response of cold acclimation. After the removal of cold, the expression patterns of most genes rapidly returned to the original states. However, photosynthetic light-harvesting complex genes and lipid metabolism-related genes stayed upregulated in cold deacclimated plants compared to non-treated plants. It is also notable that many well-known cold-inducible genes are slightly induced in reacclimation and their expression remains at relatively low levels in cold reacclimation compared to the expression during the first cold acclimation. The results in this study show the dynamic nature of gene expression occurring during cold acclimation, deacclimation and reacclimation. Our results suggest that there is a memory of cold stress and that the 'memory of cold stress' is possibly due to elevated photosynthetic efficiency, modified lipid metabolism, increased calcium signaling, pre-existing defense protein made during first cold acclimation and/or modified signal transduction from pre-existing defense protein.