Evolving bioanalytical strategies in the wake of pioneering biotherapeutics.

The development of biotherapeutics, particularly multi-domain biotherapeutics (MDBs) and antibody drug conjugates (ADCs), presents unique challenges due to their complexity and increased immunogenicity risks. Bioanalytical approaches play a pivotal role in addressing these challenges. Robust bioanalytical methods are essential for effectively assessing the pharmacokinetics (PK), pharmacodynamics (PD), and immunogenic responses of novel biologics.

Proteomic insights into broiler stress responses to LED lighting: Effects on liver proteome under neutral, cool, and warm spectra.

Light plays a critical role in poultry production, influencing broiler behavior, immune function, and growth. Effective lighting management optimizes broiler health and performance, with LED lighting emerging as an energy-efficient alternative to traditional fluorescent lights. However, the molecular impacts of different LED light spectra remain underexplored.

Unlocking the Complexity of Antibody-Drug Conjugates: A Cutting-Edge LC-HRMS Approach to Refine Drug-to-Antibody Ratio Measurements with Highly Reactive Payloads.

The complexity of therapeutic proteins like antibody-drug conjugates (ADCs) holds a tremendous analytical challenge. Complementary mass spectrometry approaches such as peptide mapping and intact mass analysis are required for the in-depth characterization of these bioconjugates. Cysteine-linked ADCs have shown a unique challenge for characterization, mainly when the conjugation is carried out on interchain cysteines, because their intact analysis requires native mass spectrometry conditions to preserve non-covalent binding between antibody chains.

Feedback from the 10th European Bioanalysis Forum Young Scientist Symposium and from the Science Café discussions on sustainability in bioanalysis.

Since 2014, the European Bioanalysis Forum has organized the Young Scientist Symposium, providing early-career scientists with unique development opportunities. By fostering a peer community, this initiative has successfully lowered barriers to engagement in cross-company, pre-competitive interactions. Since 2018, the symposium has included the Science Café roundtable, further reinforcing the European Bioanalysis Forum's commitment to supporting young scientists beyond their scientific expertise by promoting broader professional development.

Label-free liquid chromatography mass spectrometry analysis of changes in broiler liver proteins under transport stress.

Transportation duration and distance are significant concerns for animal welfare, particularly in the poultry industry. However, limited proteomic studies have investigated the impact of transport duration on poultry welfare. In this study, mass spectrometry based bottom up proteomics was employed to sensitively and impartially profile the liver tissue proteome of chickens, addressing the issue of animal stress and welfare in response to transportation before slaughter.

Label-free liquid chromatography-mass spectrometry comparison of the breast muscle proteome profiles in two fast-growing broilers.

Poultry meat-production is increasing worldwide; leading to the selection of chickens for meat-production that show a fast growth. A label-free quantitative proteomic-approach and Western-blot were applied to investigate the dynamics of muscle protein under rapid growth conditions in two common fast-growing broiler genetic-lines (Ross 508 and AZ Extra Heavy Red-chicken). Muscle exudate from chicken Pectoralis major was used as substrate to unveil the proteome of these genetic-lines.

Zinc Supplementation Improves Texture, Oxidative Stability of Caciotta Cheese and Reduces Biogenic Amines Production.

Zinc is essential for animals, playing a vital role in enzyme systems and various biochemical reactions. It is crucial to ensure a sufficient intake of zinc through the diet to maintain efficient homeostasis. Only few studies on zinc effect in cow lactating diet evaluated the effects on milk and cheese quality, with conflicting findings.

Evaluation of Chemical and Nutritional Characteristics of Ricotta Cheese from Two Different Breeds: The Endangered Italian Teramana Goat and the Cosmopolitan Saneen Goat.

The present study aimed to compare the qualitative features of ricotta cheese produced by Teramana goats and Saanen goats raised in similar breeding systems and environmental conditions. The analyses were performed on ricotta after 0 (T0) and 5 (T5) days of storage at 4 °C. Ricotta cheese samples were subjected to chemical and physical analyses.

Leveraging Cross-Linking Mass Spectrometry for Modeling Antibody-Antigen Complexes.

Elucidating antibody-antigen complexes at the atomic level is of utmost interest for understanding immune responses and designing better therapies. Cross-linking mass spectrometry (XL-MS) has emerged as a powerful tool for mapping protein-protein interactions, suggesting valuable structural insights. However, the use of XL-MS studies to enable epitope/paratope mapping of antibody-antigen complexes is still limited up to now.

Assessing MAPPs assay as a tool to predict the immunogenicity potential of protein therapeutics.

MHC-II-associated peptide proteomics (MAPPs) is a mass spectrometry-based (MS) method to identify naturally presented MHC-II-associated peptides that could elicit CD4+T cell activation. MAPPs assay is considered one of the assays that better characterize the safety of biotherapeutics by driving the selection of the best candidates concerning their immunogenicity risk. However, there is little knowledge about the impact of bead material on the recovery of MHC-II MS-eluted ligands in MAPPs assays.

Comparative Label-Free Liquid Chromatography-Mass Spectrometry Milk Proteomic Profiles Highlight Putative Differences between the Autochthon Teramana and Saanen Goat Breeds.

Goat's milk is an excellent source of nutrients, with greater benefits compared to cow's milk. Limited information is available on autochthon goat breeds, which are important for biodiversity preservation. In this study, the aim of using label-free quantification was to investigate the milk proteome of two goat breeds, the autochthon Teramana and Saanen breeds, which are commonly used by the industry.

Hybrid liquid chromatography high resolution and accuracy mass spectrometry approach for quantification of antibody-drug conjugates at the intact protein level in biological samples.

Preclinical in vivo and in vitro characterization of Antibody-Drug Conjugates (ADCs) involves the development of several bioanalytical methods to address many drug exposure questions. The current pharma industry approach requires at least three different assays that must be run, i.e.

Development of a Liquid Chromatography and High-Resolution and -Accuracy Mass Spectrometry Method to Evaluate New Biotherapeutic Entity Processing in Human Liver Lysosomes.

Biotherapeutic immunogenicity remains a great challenge for researchers because multiple factors trigger immune responses. Predicting and assessing the potential human immune response against biological drugs could represent an impressive breakthrough toward generating potentially safer and more efficacious therapeutic proteins. This article describes an in vitro assay that can contribute to evaluating the potential immunogenicity of biotherapeutics by focusing on lysosomal proteolysis.

Preclinical risk assessment strategy to mitigate the T-cell dependent immunogenicity of protein biotherapeutics: State of the art, challenges and future perspectives.

Protein therapeutics hold a prominent role and have brought significant diversity in efficacious medicinal products. Not just monoclonal antibodies and different antibody formats (pegylated antigen-binding fragments, bispecifics, antibody-drug conjugates, single chain variable fragments, nanobodies, dia-, tria- and tetrabodies), but also purified blood products, growth factors, recombinant cytokines, enzyme replacement factors, fusion proteins are all good instances of therapeutic proteins that have been developed in the past decades and approved for their value in oncology, immune-oncology, and autoimmune diseases discovery programs. Although there was an ingrained belief that fully humanized proteins were expected to have limited immunogenicity, adverse effects associated with immune responses to biological therapies raised some concern in biotech companies.

Comparative Proteomics Analysis of Pig Muscle Exudate through Label-Free Liquid Chromatography-Mass Spectrometry.

Capital-driven animal husbandry systems undertaken in the last century led to the abandoning of many pig breeds that were not profitable. These local pig breeds and their respective production systems have great potential as they are able to respond to the high criteria and needs of modern society concerning some environmental aspects, animal-welfare, healthiness, etc. This is the case of the black pigs of Italy.

New Insight into the Quality Traits of Milk and Cheese from Teramana Goats, a Native Italian Breed.

The preservation and enhancement of native breeds is a central issue to initiate new breeding policies, which are sustainable and adapted to climate changes. The aim of this study was the characterisation of the qualitative traits of milk and cheese obtained from Teramana goats compared with Saanen goats reared in the same breeding facilities or environment. The research involved 41 Teramana goats and 40 Saanen goats.

Improved Measurement of Solar Neutrinos from the Carbon-Nitrogen-Oxygen Cycle by Borexino and Its Implications for the Standard Solar Model.

We present an improved measurement of the carbon-nitrogen-oxygen (CNO) solar neutrino interaction rate at Earth obtained with the complete Borexino Phase-III dataset. The measured rate, R_{CNO}=6.7_{-0.

Salvianolic Acid B Strikes Back: New Evidence in the Modulation of Expression and Activity of Matrix Metalloproteinase 9 in MDA-MB-231 Human Breast Cancer Cells.

Salvianolic acid B (SalB) is a bioactive compound from , one of the most important traditional herbal medicines widely used in several countries for the treatment of cardiovascular diseases. The aim of this study was to evaluate the in vitro effect of SalB on the expression and the activity of matrix metalloproteinase 9 (MMP-9), a zinc-dependent proteolytic enzyme, in human MDA-MB-231 breast cancer cells. This cellular model is characterized by a marked invasive phenotype, supported by a high constitutive expression of MMPs, especially gelatinases.

A Label-Free Quantitative Analysis for the Search of Proteomic Differences between Goat Breeds.

The intensification and standardization of livestock farming are causing a decline in the number of animal breeds in many species, such as the goat. The availability of more studies on the potentiality of goat breeds could raise awareness of their importance, conservation and productive possibilities. Label-free quantitative analysis was applied in this study to investigate the proteomic differences between the autochthon Teramana and Saanen goats that could be useful for defining peculiar features of these breeds.

Label-free quantitative proteomics and stress responses in pigs-The case of short or long road transportation.

Ethical livestock production is currently a major concern for consumers. In parallel, research has shown that transport duration is an important factor affecting animal welfare and has a negative impact on the final product quality and on the production cost. This study applied proteomics methods to the animal stress/welfare problem in pigs muscle-exudate with the aim to identify proteins indicative of molecular processes underpinning transport stress and to better characterise this species as a biomedical model.