Monitoring and quantifying host cell proteins (HCPs) in biotherapeutic production processes is crucial to ensure product quality, stability, and safety. Liquid chromatography-mass spectrometry (LC-MS) analysis has emerged as an important tool for identifying and quantifying individual HCPs. However, LC-MS-based approaches face challenges due to the wide dynamic range between HCPs and the therapeutic protein as well as laborious sample preparation and long instrument time. To address these limitations, we evaluated the application of parallel accumulation-serial fragmentation combined with data-independent acquisition (diaPASEF) to HCP analysis for biopharmaceutical process development applications. We evaluated different library generation strategies and LC methods, demonstrating the suitability of these workflows for various HCP analysis needs, such as in-depth characterization and high-throughput analysis of process intermediates. Remarkably, the diaPASEF approach enabled the quantification of hundreds of HCPs that were undetectable by a standard data-dependent acquisition mode while considerably improving sample requirement, throughput, coverage, quantitative precision, and data completeness.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1021/acs.analchem.4c00977 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Morphological and molecular characteristics of swine coccidia based on single oocyst isolation.
Parasitol Res
January 2025
College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, P.R. China.
Swine coccidiosis is a host-specific protozoan disease caused by Cystoisospora suis and various Eimeria species, leading to diarrhea or subclinical signs in pigs. In this study, 3296 fecal samples from 55 farms across six provinces in China were collected and examined to determine the prevalence and molecular characteristics of swine coccidia. The single oocyst isolation technique (SOIT) and molecular characterization identified nine coccidian species, with an overall infection prevalence of 13.
View Article and Find Full Text PDFDouble-stranded RNA orbivirus disrupts the DNA-sensing cGAS-sting axis to prevent type I IFN induction.
Cell Mol Life Sci
January 2025
Centro de Investigación en Sanidad Animal, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria, Consejo Superior de Investigaciones Científicas (CISA-INIA-CSIC), Valdeolmos, Madrid, Spain.
Cyclic GMP-AMP synthase (cGAS) is a DNA sensing cellular receptor that induces IFN-I transcription in response to pathogen and host derived cytosolic DNA and can limit the replication of some RNA viruses. Some viruses have nonetheless evolved mechanisms to antagonize cGAS sensing. In this study, we evaluated the interaction between Bluetongue virus (BTV), the prototypical dsRNA virus of the Orbivirus genus and the Sedoreoviridae family, and cGAS.
View Article and Find Full Text PDFXopAE Effector from pv. Targets HSP20-Like p23 Cochaperone to Suppress Plant Basal Immunity.
Mol Plant Microbe Interact
January 2025
Universidad de los Andes, Biology, Cra 1 # 18A-10, Bogota, Cundinamarca, Colombia, 110121;
Pathogenic bacteria use Type 3 effector proteins to manipulate host defenses and alter metabolism to favor their survival and spread. The non-model bacterial pathogen pv. () causes devastating disease in cassava.
View Article and Find Full Text PDFSLO co-opts host cell glycosphingolipids to access cholesterol-rich lipid rafts for enhanced pore formation and cytotoxicity.
mBio
January 2025
Division of Infectious Diseases, Boston Children's Hospital, Boston, Massachusetts, USA.
Unlabelled: Streptolysin O (SLO) is a virulence determinant of group A (), the agent of streptococcal sore throat and severe invasive infections. SLO is a member of a family of bacterial pore-forming toxins known as cholesterol-dependent cytolysins, which require cell membrane cholesterol for pore formation. While cholesterol is essential for cytolytic activity, accumulating data suggest that cell surface glycans may also participate in the binding of SLO and other cholesterol-dependent cytolysins to host cells.
View Article and Find Full Text PDFRNASEK interacting with PEDV structural proteins facilitates virus entry via clathrin-mediated endocytosis.
J Virol
January 2025
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
Porcine epidemic diarrhea virus (PEDV), as a type of Alphacoronavirus causing acute diarrhea and high death rate among sucking piglets, poses great financial damage to the swine industry. Nevertheless, the molecular mechanism whereby PEDV enters host cells is unclear, limiting the development of PED vaccines and anti-PEDV agents. The present study found that the host protein ribonuclease kappa (RNASEK) was regulated by USF2, a transcription factor, and facilitated the PEDV replication.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!