Alphalyse’s Post

Monoclonal antibodies (mAbs) can be prone to fragmentation and clipping and in these cases, adequate removal of these species is critical during manufacturing. However, contrary to mAb aggregation, manufacturers have not given much attention to removal of fragments and clipped species and as a result most conventional mAb platforms offer at best limited capabilities for their removal. In this study, we propose a novel purification platform that uses multimodal chromatography and achieves complete removal of a range of mAb fragments and clipped products (25–120 kDa). The utility of the platform has been successfully demonstrated for 2 IgG1s and 2 IgG4s. Further, adequate removal of the various host cell impurities such as host cell proteins (<10 ppm) and host cell DNA (<5 ppb) has been achieved. Finally, the platform was able to deliver adequate removal of high molecular weight impurities (<1 %) and a 30 % clearance of the acidic charge variant. The proposed single step has been shown to deliver what typically the polishing chromatography and intermediate purification chromatography steps deliver in a traditional mAb platform. https://lnkd.in/dZ79UqEd

Having the ability to understand charge variations that may occur due to stability. Along with having complementary technologies to release assays is critical to biotherapeutics product development, manufacture ability and its overall success. Here Wu et al explores the use of microfluidic capillary electrophoresis hyphated to MS to characterize biotherapeutics charge variant charge due to force degeration conditions 🛎Overview: Force degerationnare performed so as tonunder stanf like routes of product degeration and strategies to help mitigate them 🎯Summary: Wu et al found that ZipChip-MS method was less trouble some for that traditional assay in releasee assay investigations. And found the technology particularly advantages in formulation and development. In force degeration studies Wu et al observed that under oxidative stress conditions, oxidation was found to be the major contributor to the changes in the charge variant profiles. Furhermore under pH stress conditions, Wu et al observed changes in the charge variant profile were due to increased deamidation, oxidation, and pyroglutamate formation. #massspectrometry #massspec #adc #biotherapeutics #biotech #chemistry #cancer #drugconjugate #science #biopharmaceutical #anytical #lcms #mabs #scienceandtechnology #pharmaceutical #mabs #processdevelopment #processmonitoring #qualitycontrol 908 Devices https://lnkd.in/e8v4NpKr

Happy to share another publication in the Journal of Pharmaceutical and Biomedical Analysis. In this work we describe the partial protein binding of both uracil and thymine that affect accurate DPD phenotyping. Here are a few key messages: - Recovery differences can be related to the extent of molecule protein binding - The use of cut-off levels requires extensive further optimization and harmonization - DPD phenotyping is hampered by a lack of method standardization Thank you to everyone who contributed to this work! Sylvia Roovers-Genet, Sander Streng, Maarten Broeren, Maarten Deenen, Joost van Dongen, Luc Brunsveld, Volkher Scharnhorst and Daan van de Kerkhof Check out our work entitled: 'Partial protein binding of uracil and thymine affects accurate dihydropyrimidine dehydrogenase (DPD) phenotyping' https://lnkd.in/e6fXYgi2

Here Li et al explores the use of native SEC MS for quantitative anaylis of cysteine linked ADC's 🛎Overview: Li et al methodology comprises of a first round of automatimated affinity purification. This is the followed Natie SEC MS and finaly data analysis is batchs 🎯Summery: Li et al found their quantative method was in agreement with MRM tmbased analysis. Where Li et al assay had a linear rage of 5-100ug/mL for as little as 20ul of serum. Li et al: "Our results indicate that this intact quantitation workflow can serve as an alternative generic method for high-throughput analysis, enabling an in-depth understanding of ADC stability and safety in vivo." https://lnkd.in/ezNST3BR #massspectrometry #massspec #adc #biotherapeutics #biotech #chemistry #cancer #drugconjugate #science #biopharmaceutical #anytical #lcms #mabs #scienceandtechnology #pharmaceutical #mabs #processdevelopment #processmonitoring #qualitycontrol

In this interesting paper, the authors used a synthetic biology tool to better exploit the widely used GS selection marker (split intein-mediated protein ligation of GS; SiMPl-GS). The aim was to investigate if this approach could be beneficial in cell line development - especially when trying to express recombinant proteins such as multi specific antibodies. Pools expressing mAbs, created using this system, were observed to be enriched for high producers - and the system was demonstrated to be a promising approach for expressing multi specific antibodies. #celllinedevelopment #CHO #GS #syntheticbiology #selectionmarker #multispecificantibody #bispecificantibody #biotherapeutics #biopharmaceuticals https://lnkd.in/eJuyHYtf

With so many recombinant proteins expressed in CHO cells which have been preadapted to desirable growth conditions these days, it can be easy to forget the challenges with adapting cell lines into suspension culture! Here's an interesting paper where the authors have taken a cell line engineering approach to make suspension adaptation faster and more efficient (and improve transfection efficiency) - by identifying factors that were associated with anchorage-independent growth, and then using inducible expression of these factors for suspension adaptation. #biotherapeutics #biopharmaceuticals #celllinedevelopment #cellculture #celllineengineering https://lnkd.in/eS3FkZ-s

Review Exploring FDA-Approved Frontiers: Insights into Natural and Engineered Peptide Analogues in the GLP-1, GIP, GHRH, CCK, ACTH, and α-MSH Realms by Othman Al Musaimi Biomolecules 2024, 14(3), 264; https://lnkd.in/gtCXsT_2 Abstract Peptides continue to gain significance in the pharmaceutical arena. Since the unveiling of insulin in 1921, the Food and Drug Administration (FDA) has authorised around 100 peptides for various applications. Peptides, although initially derived from endogenous sources, have evolved beyond their natural origins, exhibiting favourable therapeutic effectiveness. Medicinal chemistry has played a pivotal role in synthesising valuable natural peptide analogues, providing synthetic alternatives with therapeutic potential. Furthermore, key chemical modifications have enhanced the stability of peptides and strengthened their interactions with therapeutic targets. For instance, selective modifications have extended their half-life and lessened the frequency of their administration while maintaining the desired therapeutic action. In this review, I analyse the FDA approval of natural peptides, as well as engineered peptides for diabetes treatment, growth-hormone-releasing hormone (GHRH), cholecystokinin (CCK), adrenocorticotropic hormone (ACTH), and α-melanocyte stimulating hormone (α-MSH) peptide analogues. Attention will be paid to the structure, mode of action, developmental journey, FDA authorisation, and the adverse effects of these peptides.

Aggregation during mAbs production should be analyzed from the cultivation, fermentation and purification processes with integrated optimization

Have you ever wondered how Pepticom Ltd. overcomes the challenges that have historically made peptides poor drug candidates? We use the next generation of peptide drugs. Peptides have traditionally faced hurdles such as poor plasma stability, limited membrane permeability, and short circulation half-life, which have hindered their use in medicine. Early peptide drugs, such as insulin, were natural or close derivatives, while the second generation, from phage and mRNA display, showed modest improvements but were constrained in chemical diversity. This is where our new-generation peptides (peptidomimetics) made from macrocyclic non-natural and natural amino acids come in. This innovative approach enhances drug-like properties and extends half-lives. Our AI-driven platform integrates 600+ building blocks to explore an incredible 10^80 potential solutions, pushing the boundaries of peptide drug design. Currently, we're advancing what we believe to be the world's first AI-designed peptide for psoriasis, in our preclinical immunology project. As we recognize #PsoriasisAwarenessMonth, our lead IL-17 oral therapy targeting Plaque Psoriasis, with a market projected to reach $40B by 2029, demonstrates our commitment to advancing care and addressing unmet medical needs. Read more about how Pepticom designs de novo peptide drug candidates using proprietary AI-driven biocomputational software in Nature Magazine here >> https://lnkd.in/eJywxkeX #AI #peptides #psoriasis #drugdelivery

🎉 I am excited to share our latest findings on detecting oxidative impurities in lipid nanoparticle formulations! Ionizable lipids in mRNA-based vaccines undergo oxidative degradation into aldehydes, which form deactivated mRNA-adducts and thus require rigorous monitoring in LNP formulations. We addressed this issue by developing a nanopore electrochemical biosensor capable of massively parallel and high-throughput screening of aldehydes in LNP formulations, achieving limits of detection as low as 1.2 ppb. We believe that this work will have a significant impact on the pharmaceutical industry as an analytical tool for quality assurance in the next generation of mRNA-based therapeutics. Huge shoutout to our collaborators from Moderna, Dr. Joseph Schariter and Dr. Dipendra Gyawali, and Dr. Christopher Welch from the Indiana Consortium for Analytical Science and Engineering for their invaluable help and guidance in this project. Check out the paper below: