Enzymatic Desialylation Enables Reliable Charge Variant Characterization of Highly Glycosylated and Sialylated Fc Fusion Proteins DOI Creative Commons
Xiaona Wen, Anita P. Liu, Jing Song

et al.

ACS Pharmacology & Translational Science, Journal Year: 2025, Volume and Issue: 8(2), P. 394 - 408

Published: Jan. 23, 2025

Fusion proteins constitute a class of engineered therapeutics and have emerged as promising candidates for disease treatment. However, the structural complexity heterogeneity fusion make their characterization extremely challenging, thus, an innovative comprehensive analytical toolbox is needed. Here, first time, we demonstrate novel robust workflow to evaluate charge variants highly glycosylated protein with heavy sialylation using imaged capillary isoelectric focusing (icIEF). In development icIEF method, key factors that were systematically investigated include desialylation level, stability desialylated molecule, incubation time temperature desialylation, concentrations, urea l-arginine effects on tertiary structure, instrumental comparability. Multivariate correlation analyses subsequently applied confirm impacts parameters evaluated. Furthermore, microfluidic chip-based system coupled ultraviolet detection mass spectrometry (icIEF-UV/MS) was utilized identify critical post-translational modifications ameliorate understanding variants. Our study demonstrates this enables mechanistic heavily sialylated therapeutics.

Language: Английский

Wearable Hydrogel SERS Chip Utilizing Plasmonic Trimers for Uric Acid Analysis in Sweat DOI
Guoqun Li, Xing Zhao, Xiao Tang

et al.

Nano Letters, Journal Year: 2024, Volume and Issue: 24(42), P. 13447 - 13454

Published: Oct. 11, 2024

Uric acid is typically measured through blood tests, which can be inconvenient and uncomfortable for patients. Herein, we propose a wearable surface-enhanced Raman scattering (SERS) chip, incorporating hydrogel membrane with integrated plasmonic trimers, noninvasive monitoring of uric in sweat. The trimers feature sub 5 nm nanogaps, generating strong electromagnetic fields to boost the signal surrounding molecules. Simultaneously, pumps sweat these gaps, efficiently capturing biomarkers SERS detection. chip achieve saturation adsorption within min, eliminating variations individual production rates. Dynamic tracking lactic levels during anaerobic exercise reveals temporary suppression metabolism, likely due metabolic competition acid. Furthermore, long-term correlates well test results, confirming that regular helps reduce serum supporting its potential managing hyperuricemia.

Language: Английский

Citations

7
Monitoring Metabolic Changes in Response to Chemotherapies in Cancer with Raman Spectroscopy and Metabolomics DOI
Gabriel Cutshaw, Nora Hassan, Saji Uthaman

et al.

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(35), P. 13172 - 13184

Published: Aug. 21, 2023

Resistance to clinical therapies remains a major barrier in cancer management. There is critical need for rapid and highly sensitive diagnostic tools that enable early prediction of treatment response allow accurate decisions. Here, Raman spectroscopy was employed monitor changes key metabolites as predictors KRAS-mutant colorectal (CRC) cells, HCT116, treated with chemotherapies. We show at the single cell level HCT116 resistant cetuximab (CTX), first-line CRC, but this resistance can be overcome pre-sensitization cells oxaliplatin (OX). In combination CTX + OX, sequential delivery OX followed by rather than simultaneous administration drugs observed effective therapy. Our results demonstrated metabolic are well aligned cellular mechanical where Young's modulus decreased after treatment, indicating both properties metabolism likely responsible proliferation. findings were verified mass spectrometry (MS) metabolomics, platforms showed lipids, nucleic acids, amino acids resistance/response. Finally, pathways enriched identified when downregulated treatment. This study highlights drug-induced (Raman) ensemble have potential identify mechanisms therapies.

Language: Английский

Citations

14
Bimetallic AuNR@AgNCs for ultrasensitive surface-enhanced Raman scattering sensing of dithianon in apple juice DOI

Kaiyi Zheng,

Ye Shen, Ziyang Chen

et al.

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1292, P. 342199 - 342199

Published: Jan. 4, 2024

Language: Английский

Citations

6
A Systematic Approach toward Enabling Maximal Targeting Efficiency of Cell Surface Proteins with Actively Targeted SERS Nanoparticles DOI
Pegah Bagheri, Olga E. Eremina,

Augusta Fernando

et al.

ACS Applied Materials & Interfaces, Journal Year: 2024, Volume and Issue: 16(13), P. 15847 - 15860

Published: March 20, 2024

With their intricate design, nanoparticles (NPs) have become indispensable tools in the quest for precise cellular targeting. Among various NPs, gold NPs stand out with unique features such as chemical stability, biocompatibility, adjustable shape, and size-dependent optical properties, making them particularly promising molecular detection by leveraging surface-enhanced Raman scattering (SERS) effect. Their multiplexing abilities simultaneous identification of multiple biomarkers are important rapidly evolving landscape diverse phenotypes biomolecular profiling. However, challenge is ensuring that SERS can effectively target specific cells among cell types biomolecules high specificity. In this study, we improve functionalization optimizing targeting efficiency applications ca. 160 nm NP-based probes. Spherical conjugated antibodies epidermal growth factor receptor human 2, were incubated overexpressing these proteins, binding potential was quantified at each stage using flow cytometry to achieve optimal efficiency. We determined maintaining an average 3.5 × 105 thiols per NP, 300 18,000 cell, conducting a 15 min staining incubation 4 °C shaker, SM(PEG)12 cross-linker NP conjugation crucial highest Fluorescence imaging used parameters observe maximum ability efficiently suspended cells. These highly sensitive contrast agents demonstrate pivotal role effective active targeting, invaluable across biological environments.

Language: Английский

Citations

6
Metabolic Response to Small Molecule Therapy in Colorectal Cancer Tracked with Raman Spectroscopy and Metabolomics DOI Creative Commons
Gabriel Cutshaw, Neeraj Joshi, Xiaona Wen

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: unknown

Published: July 12, 2024

Abstract Despite numerous screening tools for colorectal cancer (CRC), 25 % of patients are diagnosed with advanced disease. Novel diagnostic technologies that early, accurate, and rapid imperative to assess the therapeutic efficacy clinical drugs identify new biomarkers treatment response. Here Raman spectroscopy (RS) was used track metabolic reprogramming in KRAS‐mutant HCT116 SW837 cells, KRAS wild‐type CC cells. RS combined multivariate analysis methods distinguished nonresponsive, partially responsive, responsive cells treated cetuximab, a monoclonal antibody EGFR inhibition, sotorasib, clinically approved inhibitor, various doses trametinib, an inhibitor MAPK pathway. Cells combination subtoxic trametinib BKM120, PI3K pathway, showed synergistic response between two pathways. Using supervised machine learning regression model, we established scoring methodology trained priori predict combinations. metabolites were verified mass spectrometry, enrichment pathways identified, including amino acid, purine, nicotinate nicotinamide metabolism differentiated monotherapy from therapy. Our approach may ultimately be applicable patient‐derived primary cultures patient tumors effective individualized care.

Language: Английский

Citations

6
Oppositely-charged silver nanoparticles enable selective SERS molecular enhancement through electrostatic interactions DOI
Yuqing Gu, Siyi Wu,

Zhewen Luo

et al.

Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy, Journal Year: 2024, Volume and Issue: 322, P. 124852 - 124852

Published: July 21, 2024

Language: Английский

Citations

6
Hypoxanthine is a metabolic biomarker for inducing GSDME-dependent pyroptosis of endothelial cells during ischemic stroke DOI Creative Commons

Jing Ye,

Xinyuan Bi, Shiyu Deng

et al.

Theranostics, Journal Year: 2024, Volume and Issue: 14(15), P. 6071 - 6087

Published: Jan. 1, 2024

Stroke induces metabolic changes in the body, and metabolites have become potential biomarkers for stroke. However, specific involved stroke mechanisms underlying brain injury during remain unclear.

Language: Английский

Citations

6
Raman Spectroscopy for Cell Analysis: Retrospect and Prospect DOI
Wenjing Xu, Wei Zhu, Y. X. Xia

et al.

Talanta, Journal Year: 2024, Volume and Issue: 285, P. 127283 - 127283

Published: Nov. 26, 2024

Language: Английский

Citations

5
Designed Nanomaterials‐Assisted Proteomics and Metabolomics Analysis for In Vitro Diagnosis DOI Open Access
Yanhui Wang, Rongxin Li, Weikang Shu

et al.

Small Methods, Journal Year: 2023, Volume and Issue: 8(1)

Published: Nov. 3, 2023

Abstract In vitro diagnosis (IVD) is pivotal in modern medicine, enabling early disease detection and treatment optimization. Omics technologies, particularly proteomics metabolomics, offer profound insights into IVD. Despite its significance, omics analyses for IVD face challenges, including low analyte concentrations the complexity of biological environments. addition, direct analysis by mass spectrometry (MS) often hampered issues like large sample volume requirements poor ionization efficiency. Through manipulating their size, surface charge, functionalization, as well nanoparticle‐fluid incubation conditions, nanomaterials have emerged a promising solution to extract biomolecules enhance desorption/ionization efficiency MS detection. This review delves last five years nanomaterial applications omics, focusing on role enrichment, separation, proteins metabolites It aims provide comprehensive update design application highlighting potential revolutionize

Language: Английский

Citations

13
A database of computed Raman spectra of inorganic compounds with accurate hybrid functionals DOI Creative Commons
Yuheng Li, D.-K. Lee, Pengfei Cai

et al.

Scientific Data, Journal Year: 2024, Volume and Issue: 11(1)

Published: Jan. 22, 2024

Abstract Raman spectroscopy is widely applied in identifying local structures materials, but the interpretation of spectra non-trivial. An accurate computational database reference calculated with a consistent level theory can significantly aid interpreting measured spectra. Here, we present inorganic compounds hybrid functionals density functional theory. were obtained by calculating dynamical matrices and polarizability tensors for from Inorganic Crystal Structure Database. The other phonon properties (e.g., infrared spectra) are stored MongoDB publicly shared through web application. We assess accuracy our calculations statistically comparing ~80 an existing experimental database. To date, contains 161 continuously growing as add more materials computed automated workflow.

Language: Английский

Citations

4