Biology subjects

Chang, K.-H.

Publications and source records attributed to Chang, K.-H..

4 recordsLinked to original sources

bioRxiv2025

A Single-Aliquot, Enrichment-Free Workflow for High-Throughput Plasma Proteome and N-Glycoproteome Profiling

High cancer mortality rates highlight an urgent need for early detection. Plasma proteomics and glycoproteomics provide minimally invasive routes for biomarker discovery, yet achieving an optimal balance among profiling depth, throughput, and longitudinal reproducibility remains a major challenge for clinical application and translation. To overcome this bottleneck, we present a single-aliquot, enrichment-free, paired-run dual-omics pipeline that concurrently profiles the global plasma proteome and N-glycoproteome from unenriched plasma. Following depletion of top14 abundant plasma proteins, we implemented sequential 23-min narrow-window data-independent acquisition (DIA) and 42-min stepped-collision-energy data-dependent (SCE-DDA) runs from the same plasma digest, delivering a clinical throughput of [~]24 patients/day with deep proteome coverage of 3,756{+/-}413 protein groups (PGs) and 1,226{+/-}78 glycopeptides per sample, including 303 FDA-approved drug targets. Cross-platform benchmarking with a previous generation instrument demonstrated significantly faster (>10-20 fold) profiling speed to achieve 113 PGs/min and high protein abundance reproducibility (Pearson r > 0.9), confirming cross-instrument transferability. Application to a 300-participant lung cohort (cancer, LDCT-detected non-cancer nodules, and controls) revealed differential expression of S100 and annexin family proteins between cancer and nodules. Paired glycoproteomic analysis (n=30) identified site-specific N-glycosylation alterations in FN1, IGHG2, C3, and MET independent of total protein abundance, uncovering additional biomarker candidates for early lung cancer detection. Together, this dual-omics strategy enables deep, scalable, and reproducible plasma analysis, supporting longitudinal biomarker discovery and validation across instruments and laboratories.

biochemistry↗

bioRxiv2022

Mesenchymal stem cell culture within perfusion bioreactors incorporating 3D-printed scaffolds enables improved extracellular vesicle yield with preserved bioactivity

Extracellular vesicles (EVs) are implicated as promising therapeutics and drug delivery vehicles in various diseases. However, successful clinical translation will depend on development of scalable biomanufacturing approaches, especially due to the documented low levels of intrinsic EV-associated cargo that may necessitate repeated doses to achieve clinical benefit in certain applications. Thus, here we assessed effects of a 3D-printed scaffold-perfusion bioreactor system on the production and bioactivity of EVs secreted from bone marrow-derived mesenchymal stem cells (MSCs), a cell type heavily implicated in generating EVs with therapeutic potential. Our results indicate that perfusion bioreactor culture results in an ~40-80-fold increase, depending on measurement method, in MSC EV production compared to conventional cell culture. Additionally, we demonstrated that MSC EVs generated using the bioreactor system significantly improved wound healing in a diabetic mouse model, with increased CD31+ staining in wound bed tissue compared to animals treated with flask cell culture-generated MSC EVs. Overall, this study establishes a promising solution to major EV translational issues (i.e., scalability and low potency) with potential for adaptation to various EV-based therapeutics and capacity for improvement alongside the continuous advancements in 3D-printing technologies.

bioengineering↗

bioRxiv2022

The Impact of Storage Condition and Duration on Function of Native and Cargo-Loaded Mesenchymal Stromal Cell Extracellular Vesicles

As evidenced by ongoing clinical trials and increased activity in the commercial sector, extracellular vesicle (EV)-based therapies have begun the transition from bench to bedside. As this progression continues, one critical aspect to EV clinical translation is understanding the effects of storage and transport conditions. Several studies have assessed the impact of storage on EV characteristics such as morphology, uptake, and component content, but effects of storage duration and temperature on EV functional bioactivity and, especially, loaded cargo are scarcely reported. Here, EV outcomes following storage at different temperatures (room temperature, 4{degrees}C, -20{degrees}C, -80{degrees}C) for various durations as well as after lyophilization were assessed. Mesenchymal stem/stromal cell (MSC) EVs were observed to retain key aspects of their bioactivity (pro-vascularization, anti-inflammation) for up to 4-6 weeks at -20{degrees}C, -80{degrees}C, and after lyophilization. Furthermore, via in vitro assays and an in vivo wound healing model respectively, these same storage conditions were also demonstrated to enable preservation of the functionality of loaded microRNA (miRNA) as well as long non-coding RNA (lncRNA) cargo in MSC EVs. These findings extend the current understanding of how EV therapeutic potential is impacted by storage conditions and may inform best practices for handling and storage of MSC EVs for both basic research and translational purposes.

bioengineering↗

bioRxiv2018

A prospective, observational study on conversion of Clinically Isolated Syndrome to Multiple Sclerosis during 4-year period (MS NEO study) in Taiwan

ImportanceCIS to MS conversion rates vary depending on population cohorts, initial manifestations, and durations of follow-up.\n\nObjectiveTo investigate conversion rate of patients from CIS to MS and the prognostic significance of demographic and clinical variables in Taiwanese population.\n\nDesignNationwide, prospective, multi-centric, observational study from November 2008 to November 2014 with 4 years follow-up.\n\nSettingMulti-centre setting at 5 institutions in Taiwan.\n\nParticipants152 patients having single clinical event potentially suggestive of MS in last 2 years were enrolled as consecutive sample. 33 patients were lost to follow-up and 16 patients did not complete the study.103 patients completed the study.\n\nIntervention(s) (for clinical trials) or Exposure(s) (for observational studies)Natural progression from first episode of CIS to MS or NMO was observed.\n\nMain Outcome(s) and Measure(s)Variables analysed were proportion of patients converting to MS or NMO after first episode of CIS, duration between first episode of neurological event and diagnosis of MS, status of anti-AQP4 IgG and length of longest contiguous spinal cord lesion in MS patients. Association between baseline characteristics and progression to MS from CIS was analyzed using multiple logistic regression. Multivariate time dependent effect of baseline characteristics on progression to MS was plotted.\n\nResults14.5% patients with CIS converted to MS after 1.1 {+/-} 1.0 years with greater predisposition (18.8%) in those having syndromes referable to the cerebral hemispheres. Conversion rate from ON to MS was 9.7%. 90.9% patients had benign disease course. 46.7% patients had abnormal MRIs at baseline, with 0.6{+/-}0.5 contrast enhanced lesions. Below normal BMI and MRI lesion load ([≥] 4 lesions) were identified as risk indicators for the development of MS. Only 4.5% were positive for anti-AQP4 antibody in MS patients and amongst them, 80% were NMO patients as diagnosed by modern criteria.\n\nConclusions and Relevance Below normal BMI and number of demyelinating lesions ([≥]4) are significant predictors of conversion from CIS to MS. A low conversion rate to MS in Taiwanese CIS patients and majority of them having a benign course and minimal disability suggest the roles of geographic, genetic and ethnic factors.\n\nTrial RegistrationNon-trial observational study.

neuroscience↗