ORCID Profile
0000-0002-1375-9820
Current Organisations
Copperbelt University
,
Mass Dynamics
,
University of South Australia
,
Hokkaido University
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Research Topics
In Research Link Australia (RLA), "Research Topics" refer to ANZSRC FOR and SEO codes. These topics are either sourced from ANZSRC FOR and SEO codes listed in researchers' related grants or generated by a large language model (LLM) based on their publications.
Top 5 Research Topics
ANZSRC Field of Research (FoR)
Proteomics and Intermolecular Interactions (excl. Medical Proteomics) | Medical Biochemistry: Proteins and Peptides (incl. Medical Proteomics) | Medical Biochemistry and Metabolomics | Analytical Biochemistry
ANZSRC Socio-Economic Objective (SEO)
Diagnostic Methods | Cancer and Related Disorders | Expanding Knowledge in the Biological Sciences |
Related Links
Publications
Magnetic enrichment of immuno-specific extracellular vesicles for mass spectrometry using biofilm-derived iron oxide nanowires
Publisher: Royal Society of Chemistry (RSC)
Date: 2023
DOI: 10.1039/D2NR05619D
Abstract: Magnetic extracellular vesicle (EV) enrichment using antibody conjugated bacteria-derived iron oxide nanowires coupled with mass spectrometry-based proteome profiling enables efficient EV subtype enrichment and reproducible proteomics.
EZYprep LC-coupled MALDI-TOF/TOF MS: An improved matrix spray application for phosphopeptide characterisation
Publisher: Wiley
Date: 07-2010
Abstract: The quality of MALDI-TOF mass spectrometric analysis is highly dependent on the matrix and its deposition strategy. Although different matrix-deposition methods have specific advantages, one major problem in the field of proteomics, particularly with respect to quantitation, is reproducibility between users or laboratories. Compounding this is the varying crystal homogeneity of matrices depending on the deposition strategy used. Here, we describe a novel optimised matrix-deposition strategy for LC-MALDI-TOF/TOF MS using an automated instrument that produces a nebulised matrix "mist" under controlled atmospheric conditions. Comparisons of this with previously reported strategies showed the method to be advantageous for the atypical matrix, 2,5-DHB, and improved phosphopeptide ionisation when compared with deposition strategies for CHCA. This optimised DHB matrix-deposition strategy with LC-MALDI-TOF/TOF MS, termed EZYprep LC, was subsequently optimised for phosphoproteome analysis and compared to LC-ESI-IT-MS and a previously reported approach for phosphotyrosine identification and characterisation. These methods were used to map phosphorylation on epidermal growth factor-stimulated epidermal growth factor receptor to gauge the sensitivity of the proposed method. EZYprep DHB LC-MALDI-TOF/TOF MS was able to identify more phosphopeptides and characterise more phosphorylation sites than the other two proteomic strategies, thus proving to be a sensitive approach for phosphoproteome analysis.
Mass spectrometry imaging as a potential tool to investigate human osteoarthritis at the tissue level
Publisher: MDPI AG
Date: 03-09-2020
DOI: 10.3390/IJMS21176414
Abstract: Osteoarthritis (OA) is the most common degenerative joint disease, predicted to increase in incidence year by year due to an ageing population. Due to the biological complexity of the disease, OA remains highly heterogeneous. Although much work has been undertaken in the past few years, underlying molecular mechanisms leading to joint tissue structural deterioration are not fully understood, with only few validated markers for disease diagnosis and progression being available. Discovery and quantitation of various OA-specific biomarkers is still largely focused on the bodily fluids which does not appear to be reliable and sensitive enough. However, with the advancement of spatial proteomic techniques, several novel peptides and proteins, as well as N-glycans, can be identified and localised in a reliable and sensitive manner. To summarise the important findings from OA biomarker studies, papers published between 2000 and 2020 were searched via Google Scholar and PubMed. Medical subject heading (MeSH) terms ‘osteoarthritis’, ‘biomarker’, ‘synovial fluid’, ‘serum’, ‘urine’, ’matrix-assisted laser desorption/ionisation’, ‘mass spectrometry imaging’, ‘proteomic’, ‘glycomic’, ‘cartilage’, ‘synovium’ AND ‘subchondral bone’ were selectively used. The literature search was restricted to full-text original research articles and written only in English. Two main areas were reviewed for OA biomarker studies: (1) an overview of disease-specific markers detected from different types of OA bio-s les, and (2) an up-to-date summary of the tissue-specific OA studies that have utilised matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI). Overall, these OA biomarkers could provide clinicians with information for better the diagnosis, and prognosis of in idual patients, and ultimately help facilitate the development of disease-modifying treatments.
Quantitative Approach Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-ToF) Mass Spectrometry
Publisher: Springer US
Date: 2021
Analysis of Phosphopeptide Changes as Spermatozoa Acquire Functional Competence in the Epididymis Demonstrates Changes in the Post-translational Modification of Izumo1
Publisher: American Chemical Society (ACS)
Date: 28-09-2012
DOI: 10.1021/PR300468M
Abstract: Spermatozoa are functionally inert when they emerge from the testes. Functional competence is conferred upon these cells during a post-testicular phase of sperm maturation in the epididymis. Remarkably, this functional transformation of epididymal spermatozoa occurs in the absence of nuclear gene transcription or protein translation. To understand the cellular mechanisms underpinning epididymal maturation, we have performed a label-free, MS-based, comparative quantification of peptides from caput, corpus and caudal epididymal spermatozoa. In total, 68 phosphopeptide changes could be detected during epididymal maturation corresponding to the identification of 22 modified proteins. Included in this list are the sodium-bicarbonate cotransporter, the sperm specific serine kinase 1, AKAP4 and protein kinase A regulatory subunit. Furthermore, four phosphopeptide changes came from Izumo1, the sperm-egg fusion protein, in the cytoplasmic segment of the protein. 2D-PAGE confirmed that Izumo1 is post-translationally modified during epididymal transit. Interestingly, phosphorylation on Izumo1 was detected on residue S339 in the caput and corpus but not caudal cells. Furthermore, Izumo1 exhibited four phosphorylated residues when spermatozoa reached the cauda, which were absent from caput cells. A model is advanced suggesting that these phospho-regulations are likely to act as a scaffold for the association of adaptor proteins with Izumo1 as these cells prepare for fertilization.
In-house packed porous graphitic carbon columns for liquid chromatography-mass spectrometry analysis of n-glycans
Publisher: Frontiers Media SA
Date: 11-06-2021
DOI: 10.3389/FCHEM.2021.653959
Abstract: Protein glycosylation is a common post-translational modification that modulates biological processes such as the immune response and protein trafficking. Altered glycosylation profiles are associated with cancer and inflammatory diseases, as well as impacting the efficacy of therapeutic monoclonal antibodies. Consisting of oligosaccharides attached to asparagine residues, enzymatically released N- linked glycans are analytically challenging due to the ersity of isomeric structures that exist. A commonly used technique for quantitative N- glycan analysis is liquid chromatography-mass spectrometry (LC-MS), which performs glycan separation and characterization. Although many reversed and normal stationary phases have been utilized for the separation of N- glycans, porous graphitic carbon (PGC) chromatography has become desirable because of its higher resolving capability, but is difficult to implement in a robust and reproducible manner. Herein, we demonstrate the analytical properties of a 15 cm fused silica capillary (75 µm i.d., 360 µm o.d.) packed in-house with Hypercarb PGC (3 µm) coupled to an Agilent 6550 Q-TOF mass spectrometer for N- glycan analysis in positive ion mode. In repeatability and intermediate precision measurements conducted on released N- glycans from a glycoprotein standard mixture, the majority of N- glycans reported low coefficients of variation with respect to retention times (≤4.2%) and peak areas (≤14.4%). N- glycans released from complex s les were also examined by PGC LC-MS. A total of 120 N- glycan structural and compositional isomers were obtained from formalin-fixed paraffin-embedded ovarian cancer tissue sections. Finally, a comparison between early- and late-stage formalin-fixed paraffin-embedded ovarian cancer tissues revealed qualitative changes in the α2,3- and α2,6-sialic acid linkage of a fucosylated bi-antennary complex N- glycan. Although the α2,3-linkage was predominant in late-stage ovarian cancer, the alternate α2,6-linkage was more prevalent in early-stage ovarian cancer. This study establishes the utility of in-house packed PGC columns for the robust and reproducible LC-MS analysis of N- glycans.
MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney
Publisher: Springer Science and Business Media LLC
Date: 02-12-2015
Design and Control of a Grid-connected Seven Level Inverter for Photovoltaic Applications
Publisher: ICT Association of Zambia
Date: 07-03-2019
DOI: 10.33260/ZICTJOURNAL.V3I1.72
Abstract: This paper presents the design and closed-loop current control of a grid connected seven-level, 3-phase diode-cl ed multilevel inverter for Photovoltaic (PV) applications. The proposed closed loop current control technique is based on the voltage-oriented proportional integral (PI) controller theory. The modulation technique used is level-shifted-carrier sinusoidal pulse width modulation (SPWM). The gain values of PI controller were selected to achieve good current quality and dynamic response. Grid synchronization was achieved by using a synchronous-reference frame phase-locked loop (SRF-PLL). Matlab/Simulink was used for the control system design and simulation. The simulation results show that a 1.34% total harmonic distortion (THD) of the output current was achieved which is within the allowable current distortion limits by international standards. The stability of the system was analyzed using pole-zero mapping and root locus.
Raw N-glycan mass spectrometry imaging data on formalin-fixed mouse kidney
Publisher: Elsevier BV
Date: 12-2018
Rapid separation and identification of beer spoilage bacteria by inertial microfluidics and MALDI-TOF mass spectrometry
Publisher: Royal Society of Chemistry (RSC)
Date: 2019
DOI: 10.1039/C9LC00152B
Abstract: Microfluidics and MALDI-TOF MS is a rapid, high-throughput, and accurate method for the identification of beer spoilage bacteria.
Imatinib as a potential antiresorptive therapy for bone disease
Publisher: American Society of Hematology
Date: 06-2006
DOI: 10.1182/BLOOD-2005-09-3568
Abstract: Osteoclasts (OCs) are large multinucleated cells derived from progenitor cells of the monocyte-macrophage lineage. Signal transduction via the macrophage–colony-stimulating factor (M-CSF) receptor, c-fms, is essential for OC formation. Since we have previously demonstrated inhibition of c-fms by imatinib, we examined the effect of imatinib on OC formation and activity. OC formation was not affected by concentrations of 1.0 μM imatinib and lower, but was reduced by 75% at 3.0 μM imatinib. In contrast, both the area of resorption and the number of resorption lacunae were reduced by 80% at 0.3 μM imatinib, and no resorption was observed at concentrations above 3.0 μM. A dose-dependent decrease in receptor activator of nuclear factor κB (RANK) expression was observed in OCs when cultured in the presence of imatinib, providing a mechanism for the decrease in OC function. In vivo analysis of the effect of imatinib on OC activity in adult mice following 8 weeks of imatinib treatment also demonstrated a decrease in OC activity. These results suggest that imatinib may have therapeutic value as an antiosteolytic agent in diseases such as osteoporosis, metastatic bone disease, and multiple myeloma.
Cancer Tissue Classification Using Supervised Machine Learning Applied to MALDI Mass Spectrometry Imaging
Publisher: MDPI AG
Date: 27-10-2021
Abstract: Matrix assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) can determine the spatial distribution of analytes such as protein distributions in a tissue section according to their mass-to-charge ratio. Here, we explored the clinical potential of machine learning (ML) applied to MALDI MSI data for cancer diagnostic classification using tissue microarrays (TMAs) on 302 colorectal (CRC) and 257 endometrial cancer (EC)) patients. ML based on deep neural networks discriminated colorectal tumour from normal tissue with an overall accuracy of 98% in balanced cross-validation (98.2% sensitivity and 98.6% specificity). Moreover, our machine learning approach predicted the presence of lymph node metastasis (LNM) for primary tumours of EC with an accuracy of 80% (90% sensitivity and 69% specificity). Our results demonstrate the capability of MALDI MSI for complementing classic histopathological examination for cancer diagnostic applications.
Mapping the testicular interstitial fluid proteome from normal rats
Publisher: Wiley
Date: 09-2016
Abstract: Communication between the testicular somatic (Sertoli, Leydig, peritubular myoid, macrophage) and germ cell types is essential for sperm production (spermatogenesis), but the communicating factors are poorly understood. We reasoned that identification of proteins in the testicular interstitial fluid (TIF) that bathes these cells could provide a new means to explore spermatogenic function. The aim of this study was to map the proteome of TIF from normal adult rats. Low-abundance proteins in TIF were enriched using ProteoMiner beads and identified by MALDI-MS/MS, recognizing 276 proteins. Comparison with proteomic and genomic databases showed these proteins originated from germ cells, somatic cells (Sertoli, peritubular myoid, Leydig), and blood plasma. In silico analysis revealed homologues of >80% TIF proteins in the human plasma proteome, suggesting ready exchange between these fluids. Only 36% of TIF proteins were common with seminiferous tubule fluid that transports mature spermatids to the epididymis, indicating these two fluids are quite different. This TIF proteome provides an important new resource for the study of intercellular communication in the testis.
Mass Dynamics 2.0: An improved modular web-based platform for accelerated proteomics insight generation and decision making
Publisher: Cold Spring Harbor Laboratory
Date: 14-12-2022
DOI: 10.1101/2022.12.12.517480
Abstract: Data processing is essential to reliably generate knowledge from proteomics studies. The complexity of the proteomics data, as well as the ability of research teams to adopt complex analysis pipelines, have proven to be an obstacle to effective collaboration and more efficient biological insight generation. Here, we introduce MD 2.0, a cloud- and web-based platform for quantitative proteomics data, which implements a novel analysis workspace where statistical analyses, visualizations, and external knowledge generation are integrated into a modular framework. This modularity enables researchers the flexibility to test different hypotheses, and customize and template complex proteomics analyses, thereby expediting insight generation for complex datasets. The extensible MD 2.0 environment has been built with a scalable architecture to allow rapid development of future analysis modules and enhanced tools for remote collaboration, like experiment sharing and a live chat capability. The new drag-and-drop modules allow researchers to easily and quickly assess different aspects of an experiment, including quality control, differential expression and enrichment analysis. The modularity of MD 2.0 lays the foundation to support broader community-based analytical template generation and optimized sharing and collaboration between proteomics experts and biologists, thereby accelerating research teams’ abilities to extract knowledge from complex proteomics datasets.
Tyrosine phosphorylation enrichment and subsequent analysis by MALDI-TOF/TOF MS/MS and LC-ESI-IT-MS/MS
Publisher: Wiley
Date: 11-2010
DOI: 10.1002/0471140864.PS1311S62
Abstract: This unit describes methods to detect, identify, and characterize tyrosine phosphorylation in proteins by mass spectrometry, including s le preparation methods, enrichment strategies using phosphotyrosine‐specific antibodies, and chromatographic separation methods. Curr. Protoc. Protein Sci . 62:13.11.1‐13.11.26. © 2010 by John Wiley & Sons, Inc.
Reliability assessment in active distribution networks with detailed effects of PV systems
Publisher: Springer Science and Business Media LLC
Date: 03-2014
MALDI mass spectrometry imaging of early- and late-stage serous ovarian cancer tissue reveals stage-specific N-glycans
Publisher: Wiley
Date: 16-08-2019
Abstract: Epithelial ovarian cancer is one of the most fatal gynecological malignancies in adult women. As studies on protein N-glycosylation have extensively reported aberrant patterns in the ovarian cancer tumor microenvironment, obtaining spatial information will uncover tumor-specific N-glycan alterations in ovarian cancer development and progression. matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) is employed to investigate N-glycan distribution on formalin-fixed paraffin-embedded ovarian cancer tissue sections from early- and late-stage patients. Tumor-specific N-glycans are identified and structurally characterized by porous graphitized carbon-liquid chromatography-electrospray ionization-tandem mass spectrometry (PGC-LC-ESI-MS/MS), and then assigned to high-resolution images obtained from MALDI-MSI. Spatial distribution of 14 N-glycans is obtained by MALDI-MSI and 42 N-glycans (including structural and compositional isomers) identified and structurally characterized by LC-MS. The spatial distribution of oligomannose, complex neutral, bisecting, and sialylated N-glycan families are localized to the tumor regions of late-stage ovarian cancer patients relative to early-stage patients. Potential N-glycan diagnostic markers that emerge include the oligomannose structure, (Hex)
A sensitive magnetic bead method for the detection and identification of tyrosine phosphorylation in proteins by MALDI-TOF/TOF MS
Publisher: Wiley
Date: 06-2009
Abstract: Phosphorylation is one of the most important PTMs and is estimated to occur on 30% of the mammalian proteome. Its perturbed regulation has been implicated in many pathologies. The rarity of phosphotyrosine compared with phosphoserine or phosphothreonine is prompting the development of more sensitive approaches because proteomic technologies that are currently used to assess tyrosine phosphorylation in proteins are inadequate, identifying only a fraction of the predicted tyrosine phosphoproteome. Here we describe the development of a reproducible, high-sensitivity methodology for the detection and mapping of phosphotyrosine residues by MS. The anti-phosphotyrosine antibody 4G10 was coupled covalently to super para-magnetic beads or by affinity to super para-magnetic beads with protein G covalently attached. Using this approach, we successfully enriched phosphotyrosine peptides mixed with non-phosphorylated peptides at a ratio of up to 1:200, enabling detection at a level representing the highest sensitivity reported for tyrosine phosphorylation. The beads were subsequently used to enrich tyrosine phosphopeptides from a digest of the in vitro-phosphorylated recombinant beta-intracellular region of the granulocyte-macrophage colony-stimulating factor receptor, which was subsequently analysed by MALDI-TOF/TOF MS. Our results define this methodology as a sensitive approach for tyrosine phosphoproteome analysis.
Increased Phospho-Keratin 8 Isoforms in Colorectal Tumors Associated with EGFR Pathway Activation and Reduced Apoptosis
Publisher: Hindawi Limited
Date: 31-01-2012
DOI: 10.5402/2012/706545
Abstract: Hyperphosphorylated keratin (K) 8 acts as a phosphate “sponge” for stress-activated protein kinases thereby inhibiting pro-apoptotic molecules and thus apoptosis. MAP kinase/ERK1 has increased activity in colorectal cancer (CRC) and is known to phosphorylate K8. The aims were to identify the K8 isoforms abundantly present in colon tumors, using 2D difference gel electrophoresis (DIGE), to identify the modifications using mass spectrometry, and to validate the differential abundance of these isoforms in tumors relative to matched normal mucosae. 2D DIGE showed 3 isoforms of K8 significantly increased in tumor ≥2-fold in 6/8 pairs. Metal oxide affinity chromatography mass spectrometry and bioinformatics were used to identify phosphorylated serine residues. Levels of PS24, PS432, and PS74 by western blotting were found to be significantly increased in tumor versus matched normal. Blocking of EGFR signaling in Caco2 cells showed a significant decrease ( P 0.0001 ) in K8 PS74 and PS432 levels by 59% and 66%, respectively, resulting in increased apoptosis.
Quantitation and Identification of Intact Major Milk Proteins for High-Throughput LC-ESI-Q-TOF MS Analyses
Publisher: Public Library of Science (PLoS)
Date: 17-10-2016
Quantitative mass spectrometry-based analysis of proteins related to cattle and their products – focus on cows' milk beta-casein variants
Publisher: Elsevier BV
Date: 02-2021
Balancing sufficiency and impact in reporting standards for mass spectrometry imaging experiments
Publisher: Oxford University Press (OUP)
Date: 14-08-2018
Translating N-glycan analytical applications into clinical strategies for ovarian cancer
Publisher: Wiley
Date: 09-11-2019
Abstract: Protein glycosylation, particularly N-linked glycosylation, is a complex posttranslational modification (PTM), which plays an important role in protein folding and conformation, regulating protein stability and activity, cell-cell interaction, and cell signaling pathways. This review focuses on analytical techniques, primarily MS-based techniques, to qualitatively and quantitatively assess N-glycosylation while successfully characterizing compositional, structural, and linkage features with high specificity and sensitivity. The analytical techniques explored in this review include LC-ESI-MS/MS and MALDI time-of-flight MS (MALDI-TOF-MS), which have been used to analyze clinical s les, such as serum, plasma, ascites, and tissue. Targeting the aberrant N-glycosylation patterns observed in MALDI-MS imaging (MSI) offers a platform to visualize N-glycans in tissue-specific regions. The studies on the intra-patient (i.e., a comparison of tissue-specific regions from the same patient) and inter-patient (i.e., a comparison of tissue-specific regions between different patients) variation of early- and late-stage ovarian cancer (OC) patients identify specific N-glycan differences that improve understanding of the tumor microenvironment and potentially improve therapeutic strategies for the clinic.
An Efficient Hybrid Particle Swarm and Gradient Descent Method for the Estimation of the Hosting Capacity of Photovoltaics by Distribution Networks
Publisher: MDPI AG
Date: 06-07-2023
DOI: 10.3390/EN16135207
Abstract: With many distribution networks adopting photovoltaic (PV) generation systems in their networks, there is a significant risk of over-voltages, reverse power flow, line congestion, and increased harmonics. Therefore, there is a need to estimate the amount of PV that can be injected into the distribution network without pushing the network towards these threats. The largest amount of PV a distribution system can accommodate is the PV hosting capacity (PVHC). The paper proposes an efficient method for estimating the PVHC of distribution networks that combines particle swarm optimization (PSO) and the gradient descent algorithm (GD). PSO has a powerful exploration of the solution space but poor exploitation of the local search. On the other hand, GD has great exploitation of local search to obtain local optima but needs better global search capabilities. The proposed method aims to harness the advantages of both PSO and GD while alleviating the ills of each. The numerical case studies show that the proposed method is more efficient, stable, and superior to the other meta-heuristic approaches.
Use of Titanium Dioxide To Find Phosphopeptide and Total Protein Changes During Epididymal Sperm Maturation
Publisher: American Chemical Society (ACS)
Date: 27-01-2011
DOI: 10.1021/PR1007224
Abstract: Although the overall performance of modern mass spectrometers has increased, proteomic analysis of complex s les still requires prefractionation either at the protein or peptide level to allow for in-depth analysis of normal cellular function. Here, we report a novel way to identify protein changes occurring during sperm development through the epididymis. Phosphopeptides were first enriched from either the rat caput or caudal regions of the epididymides using TiO(2), and the profiles then quantitatively compared. We show that 77 TiO(2)-enriched peptides become significantly modified in the epididymis, equating to 53 proteins. Through the use of immunoblot analysis, we confirmed that three proteins, ornithine-decarboxylase antizyme 3, heat-shock protein 90α, and testis-lipid binding protein, undergo major protein loss during epididymal passage. Many other proteins, including t-complex protein 10 and Spata18 show testis unique expression, appear to undergo phosphorylation during this same time frame. These data provide mechanistic insight into the means by which spermatozoa acquire functionality during epididymal transit.
Gelatin-coated indium tin oxide slides improve human cartilage-bone tissue adherence and N-glycan signal intensity for mass spectrometry imaging
Publisher: Springer Science and Business Media LLC
Date: 15-10-2021
The utility of isotope-coded protein labeling for prioritization of proteins found in ovarian cancer patient urine
Publisher: American Chemical Society (ACS)
Date: 16-08-2013
DOI: 10.1021/PR400618V
Abstract: Urine offers a number of attractive features as a s le type for biomarker discovery, including noninvasive s ling, quantity and availability, stability, and a narrow dynamic range. In this study we report the first application of isotope coded protein labeling (ICPL), coupled with in-solution isoelectric fractionation and LC-MALDI-TOF/TOF, to examine and prioritize urinary proteins from ovarian cancer patients. Following the definition of stringent exclusion criteria a total of 579 proteins were identified with 43% providing quantitation data. Protein abundance changes were validated for selected proteins by ESI-Qq-TOF MS, following which Western blot and immunohistochemical analysis by tissue microarray was used to explore the biological relevance of the proteins identified. Several established markers (e.g., HE4, osteopontin) were identified at increased levels in ovarian cancer patient urine, validating the approach used we also identified a number of potential marker candidates (e.g., phosphatidylethanolamine binding protein 1, cell-adhesion molecule 1) previously unreported in the context of ovarian cancer. We conclude that the ICPL strategy for identification and relative quantitation of urine proteins is an appropriate tool for biomarker discovery studies, and can be applied for the selection of potential biomarker candidates for further characterization.
Strategy for Automated Development of
Reversed‐Phase HPLC
Methods for
Domain‐Specific
Characterization of Monoclonal Antibodies
Publisher: Wiley
Date: 06-08-2021
Scavenging of soluble and immobilized CCL21 by ACKR4 regulates peripheral dendritic cell emigration
Publisher: Proceedings of the National Academy of Sciences
Date: 19-04-2021
Abstract: The immune system relies on coordinated interactions between motile cells guided by molecules known as chemokines. However, processes that control chemokine distribution in complex in vivo microenvironments are poorly understood. Dendritic cells in barrier tissues require the chemokine CCL21 to enter lymphatic vessels during tissue egress. Here, we demonstrate that ACKR4 shapes CCL21 distribution in barrier tissues and prevents leakage of CCL21 from the tissue. Without ACKR4, extracellular CCL21 gradients in barrier sites are saturated and nonfunctional, DCs cannot home directly to lymphatic vessels, and excess soluble CCL21 from peripheral tissues pollutes lymph nodes. The results increase understanding of regulation of dendritic cell egress and chemokine distribution in vivo and raise new questions regarding the function of solubilized CCL21.
Egg White as a Quality Control in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI-MSI)
Publisher: American Chemical Society (ACS)
Date: 29-10-2019
DOI: 10.1021/ACS.ANALCHEM.9B03091
Abstract: The strength of MALDI-MSI is to analyze and visualize spatial intensities of molecular features from an intact tissue. The distribution of the intensities can then be visualized within a single tissue section or compared in between sections, acquired consecutively. This method can be reliably used to reveal physiological structures and has the potential to identify molecular details, which correlate with biological outcomes. MALDI-MSI implementation in clinical laboratories requires the ability to ensure method quality and validation to meet diagnostic expectations. To be able to get consistent qualitative and quantitative results, standardized s le preparation and data acquisition are of highest priority. We have previously shown that the deposition of internal standards onto the tissue section during s le preparation can be used to improve the mass accuracy of monitored
An external quality assurance trial to assess mass spectrometry protein testing facilities for identifying multiple human peptides
Publisher: Springer Science and Business Media LLC
Date: 05-08-2019
DOI: 10.1007/S00216-019-02047-Y
Abstract: The application of proteomic liquid chromatography mass spectrometry (LC-MS) for identifying proteins and peptides associated with human disease is rapidly growing in clinical diagnostics. However, the ability to accurately and consistently detect disease-associated peptides remains clinically uncertain. Variability in diagnostic testing occurs in part due to the absence of appropriate reference testing materials and standardised clinical guidelines for proteomic testing. In addition, multiple proteomic testing pipelines have not been fully assessed through external quality assurance (EQA). This trial was therefore devised to evaluate the performance of a small number of mass spectrometry (MS) testing facilities to (i) evaluate the EQA material for potential usage in a proteomic quality assurance program, and to (ii) identify key problem areas associated with human peptide testing. Five laboratories were sent six peptide reference testing s les formulated to contain a total of 35 peptides in differing ratios of light (natural) to heavy (labelled) peptides. Proficiency assessment of laboratory data used a modified approach to similarity and dissimilarity testing that was based on Bray-Curtis and Sorensen indices. Proficiency EQA concordant consensus values could not be derived from the assessed data since none of the laboratories correctly identified all reference testing peptides in all s les. However, the produced data may be reflective of specific inter-laboratory differences for detecting multiple peptides since no two testing pipelines used were the same for any laboratory. In addition, laboratory feedback indicated that peptide filtering of the reference material was a common key problem area prior to analysis. These data highlight the importance of an EQA programme for identifying underlying testing issues so that improvements can be made and confidence for clinical diagnostic analysis can be attained.
A flexible stochastic PV hosting capacity framework considering network over-voltage tolerance
Publisher: Elsevier BV
Date: 03-2023
Magnetic Enrichment of Immuno-Specific Extracellular Vesicles for Mass Spectrometry Using Biofilm-Derived Iron Oxide Nanowires
Publisher: Cold Spring Harbor Laboratory
Date: 05-2022
DOI: 10.1101/2022.05.01.490183
Abstract: Immuno-specific enrichment of extracellular vesicles (EVs) originating from specific cells/tissues is a promising source of information towards improving insights into cellular pathways underpinning various pathologies and developing novel non-invasive diagnostic methods. Enrichment is an important aspect in mass spectrometry-based analyses of EVs. Herein, we report a protocol for immuno-magnetic enrichment of subtype specific EVs and their subsequent processing for mass spectrometry. Specifically, we conjugated placental alkaline phosphatase (PLAP) antibodies to magnetic iron oxide nanowires (NWs) derived from bacterial biofilms and demonstrated the utility of this approach by enriching placental specific EVs (containing PLAP) from cell culture media. We demonstrate efficient PLAP+ve EV enrichment for both NW-PLAP and Dynabeads™-PLAP, with PLAP protein recovery (83.7±8.9% and 83.2±5.9%, respectively), high particle-to-protein ratio (7.5±0.7×10 9 and 7.1 ± 1.2×10 9, respectively), and low non-specific binding of non-target EVs (7±3.2% and 5.4±2.2%, respectively). Furthermore, our optimized EV enrichment and processing approach identified 2518 and 2545 protein groups with mass spectrometry for NW-PLAP and Dynabead™-PLAP, respectively, with excellent reproducibility (Pearson correlation 0.986 and 0.988). The proposed immuno-specific EVs enrichment and liquid chromatography-tandem mass spectrometry method using naturally occurring iron oxide magnetic NWs or gold-standard Dynabeads™ enables high-quality EV proteomic studies.
Related Organisations
Related Funding Activities
Linkage Projects - Grant ID: LP110100693
Start Date: 09-2011
End Date: 12-2013
Amount: $180,000.00
Funder: Australian Research Council
View Funded Activity