This study discusses the advanced software-supported Top-Down characterization of biopharmaceutical proteins, addressing the limitations of traditional bottom-up analyses. Despite historical challenges in Top-Down sequence analysis due to inadequate software and instrumentation, this research presents a comprehensive method using modern software tools and fragmentation techniques for high...
IgGs mediate diverse immunological responses by binding antigens via their Fab and interacting with downstream receptors (e.g., Fcγ) via their Fc region, activating different pathways. Their remarkable variability originates from the B cell differentiation and activation in response to antigens, resulting in an immense repertoire of different antigen-specific clones. In addition, the immune...
Liquid chromatography is a vital analytical technique widely employed in pharmaceuticals, biotechnology, food and beverage industries, and environmental monitoring. This presentation will explore sustainable alternatives to traditional reversed-phase chromatography, which accounts for over 75% of applications and typically relies on environmentally harmful solvents like acetonitrile.
The role...
Neurodegenerative diseases are one of the most common disorders in the world, and despite intensive research, the understanding of these diseases is limited. Alzheimer's disease is the most common neurodegenerative disease, affecting about 50 million people worldwide. In addition to amyloid plaques composed of amyloid-β, neurofibrillary tangles formed from the protein Tau are a hallmark of...
Top-down proteomics is a powerful tool for investigating complex biological processes in living organisms, both in health and disease. Unlike the more common bottom-up approach, top-down proteomics does not involve digesting proteins into peptides, providing a more comprehensive view of the proteome landscape. However, using intact proteins instead of peptides presents experimental challenges...
False Discovery Rate (FDR) estimation is critical in proteomics to minimize false positive (FP) identifications. Traditional FDR estimation via the Target-Decoy Approach (TDA) mainly accounts for FPs due to fragment mass coincidences but overlooks those introduced by errors in precursor mass determination. In top-down proteomics (TDP), where spectral deconvolution assigns precursor masses,...
Bacterial infections are among the leading causes of morbidity and mortality worldwide, posing a significant threat to public health. The rapid emergence of antimicrobial resistance has further exacerbated the challenges in treating infections, making timely and accurate diagnosis crucial for effective patient management. Current methods for bacterial infection diagnosis typically involve two...
Top-down proteomics has emerged as a powerful analytical approach for characterizing intact proteins and their proteoforms offering unique insights into protein structure, post-translational modifications (PTMs), and biological function. Proteoform identification is generally achieved by separation of intact proteins using reversed-phase liquid chromatography (LC) followed by fragmentation and...
The prevalence of diabetes is steadily increasing, prompting the widespread use of various commercially available insulin analogs products in clinical practice. Assessing the chemical and structural stability of these pharmaceuticals should be carefully addressed, as their long-term efficacy and safety are intrinsically linked to chemical degradation pathways, such as deamidation [1]....
Immunoglobulins are a class of proteins that are crucial for the immune response to fight diseases. They contain many different post-translational modifications, and in particular, changes in the glycosylation patterns of immunoglobulins subclass-G (IgGs) can be linked to disease progression. In our project we aim to develop capillary-based LC-MS methods to characterize intact IgG secreted by...
Protein characterization is a very wide-reaching discipline which can encompass a broad range of techniques and approaches. Mass spectrometry is making an increasingly significant contribution to this field as the technology matures and top-down workflows are improved upon with new hardware and software innovations. The new timsOmniTM platform combines trapped ion mobility spectrometry (TIMS)...
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a single-stranded RNA virus that caused an outbreak of coronavirus disease in 2019 (COVID-19). The replication of the viral genome is facilitated by the replication/transcription complex (RTC), in which nonstructural proteins (nsps) are the principal contributors. Therefore, unraveling the functional properties and structural...
The microtubule binding protein Tau (MAPT) has been implicated as a driver of diverse progressive neurodegenerative diseases such as Alzheimer’s disease. Six splicing isoforms of the Tau protein and over 70 different post-translational modifications (PTMs) have been identified on Tau that are believed to impact fibril formation and ultimately disease progression. Though the general prevalence...
Recombinant AAV (rAAV) vectors are a leading gene delivery system due to their low immunogenicity, tissue specificity, and high efficiency. Monitoring intact AAV capsid proteins (VPs) and their post-translational modifications (PTMs) is critical for ensuring product quality and efficacy. A crucial PTM to monitor is deamidation as it can alter product potency, stability, and function1....
