Raphael Nir

Application of CGE in cell and gene therapy https://lnkd.in/eARwXTx3 Tuesday October 22nd 2024 08:00 PDT / 11:00 EDT / 16:00 BST / 17:00 CEST Capillary electrophoresis can be used for a variety of analytical assays to monitor critical quality attributes (CQAs) within the cell and gene therapy field. Plasmids, used as a critical starting material in the manufacture of AAV, require high degree of supercoiling for increased transfection efficiency resulting in increased AAV yields, therefore it is important to monitor plasmid isoform distribution for purity determination. Purity of AAV itself is also a necessary CQA to demonstrate appropriate viral protein expression and expected VP ratio, a key element for correct capsid formation. It is also important to monitor the encapsulated genome to verify that the expected genome size has been packaged. The variety of plasmids required for AAV production can lead to challenging purity analysis based on differences in size and structure. Therefore, the new DNA 20 kb Plasmid and Linear kit from SCIEX was assessed to address this challenge. Using Pharmaron’s extensive analytical method development experience, here they present an overview of innovative platform capillary gel electrophoresis methods which have been developed to assess AAV and a variety of plasmid sizes. What are plasmids and how are they used for viral vector manufacturing How can quantitative plasmid purity analysis for a variety of constructs be streamlined What information does CGE provide about viral vector quality attributes How have developments in technology allowed for high throughput cell and gene therapy analysis

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HORIZON 1.2.0 Features four foundational modules to empower your lab with genomic medicine insights from whole genome data. HORIZON enables: 💡 Rare diseases and Pan Cancer germline variant classification to enable genetic diagnosis 💡 Tumor-germline pair (tumor or liquid biopsy) vcf analysis to detect accurate somatic mutations 💡 Trio analysis for rare disease inherited and de novo variant analysis. 💡 All variant classification analysis (SNV/Indel/CNV) in one intuitive platform 💡 Ultrafast vcf analysis and automation in reporting 💡 Validated prediction in variant classification and evidence from sceintific literature Please book a demo for more information regarding HORIZON! https://lnkd.in/dFdgNzXS #cancer #rarediseases #genomics #liquidbiopsy #somaticmutation #genome

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From cell isolation and nucleic acid amplification to complete NGS workflows, ResolveServices℠ from BioSkryb make it easier than ever to start single-cell analysis. Get the most comprehensive view of the genome, transcriptome and targeted proteins, with support tailored to your project’s specific needs. Resolve More with ResolveServices: https://lnkd.in/emA8qnmg #multiomics #singlecell #genomics

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From MBoC The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis and maintain translational repression of maternal mRNA and oocyte quality The mechanisms regulating phase transitions of oogenic RNA-binding proteins are critical for oocyte quality. Schisa et. al identify the CCT chaperonin and actin as novel regulators of ER structure, RNA-binding protein phase transitions, and translational regulation in C. elegans oocytes. https://lnkd.in/eSAgmfa8

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Imagine disease-relevant cell models with unmatched precision and knockout efficiencies of ~90% - That’s the power of iPS cells + CRISPR genome editing! In our latest Tech Note, we walk you through how you can leverage induced pluripotent stem (iPS) cells combined with CRISPR technology to accelerate your discoveries. Download now and see the difference: https://hubs.la/Q02_scb50 #CRIPSR #iPScells #genomeediting #geneediting #diseasemodel

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Dual-Targeting CD38 and BCMA CAR-T Cell Immunotherapy The dual-targeted CD38 and BCMA CAR-T cell immunotherapy developed by our team is an innovative approach to treat multiple myeloma and other hematological malignancies. By simultaneously targeting CD38 and BCMA, the therapy aims to improve treatment efficacy and overcome the limitations associated with single-target CAR-T cells, especially for patients who develop resistance. Both CD38 and BCMA are highly expressed on multiple myeloma cells, making them ideal targets. While BCMA is primarily found in plasma cells, CD38 is expressed in a variety of hematological malignancies, making it more widely applicable. This dual-targeted CAR-T cell immunotherapy works by equipping T cells with chimeric antigen receptors (CARs) that recognize CD38 and BCMA antigens, enabling simultaneous or sequential targeting of both proteins on malignant cells. This not only enhances tumor cell recognition, but also increases the likelihood of tumor cell elimination and reduces the risk of antigen escape, a common problem with single-target therapies. In general, antigen escape occurs when tumor cells lose or downregulate the targeted antigen, leading to treatment resistance. By targeting two antigens, dual-targeted CAR-T cell immunotherapy provides built-in redundancy, thereby improving its efficacy. In addition, dual-targeted CAR-T cells are designed to improve persistence in patients, thereby achieving long-term immune surveillance and reducing relapse rates. However, dual-targeted therapies also have potential challenges, including increased risk of targeted off-tumor toxicity. For example, CD38 is present in some normal cells, and over-targeting may damage healthy tissues. To this end, we are using advanced engineering technologies to enhance specificity and minimize toxicity, such as combining dual signal activation to ensure that CAR-T cells are only activated in the presence of two antigens. Overall, the dual-targeted CD38 and BCMA CAR-T immunotherapy developed by our team provides a promising and powerful new option for the treatment of resistant hematological cancers, and clinical trials are currently underway to optimize its safety and efficacy. References [1] Heng Mei et al., J Hematol Oncol 2021 (doi: 10.1186/s13045-021-01170-7) [2] Yuanyan Tang et al., J Exp Clin Cancer Res 2022 (doi: 10.1186/s13046-021-02214-z) #CAR_Therapy #DualTargeting #CD38 #BCMA #MultipleMyeloma #CancerImmunotherapy #HematologicCancers #BiotechNews #CellTherapy #CancerResearch

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ਇਕ ਥਰੈਪੀ ਆ ਜੋ ਕੈਂਸਰ ਦੇ ਇਲਾਜ ਲਈ ਅਚੂਕ ਮੰਨੀ ਗਈ ਹੈ ... ਇਸ ਉਪਰ ਖਰਚਾ ਵੀ ਇਕ ਕਰੋੜ ਦੇ ਕਰੀਬ ਆਉਂਦਾ ਹੈ ... ਇਸ ਦੇ ਮੁਕਾਬਲੇ ਮੇਰੇ ਦਿਮਾਗ ਵਿੱਚ mRNA ਥਰੈਪੀ ਆਉਂਦੀ ਹੈ ਜਿਸ ਉਪਰ ਖਰਚਾ ਬਹੁਤ ਘਟ ਆਉਂਦਾ ਹੈ ਪਰ ਦੇਖਿਆ ਗਿਆ ਕਿ ਇਲਾਜ ਕਰਵਾਉਣ ਵਾਲੇ ਜੇ Afford ਕਰ ਸਕਦੇ ਹੋਣ ਤਾਂ CART-CELL Therapy ਉਪਰ ਕਰੋੜ ਰੁਪਿਆ ਇਜਰਾਇਲ ਜਾ ਕੇ ਖਰਚਣ ਨੂੰ prefer ਕਰਦੇ ਹਨ ਆਓ ਜਾਣੀਏ ਇਹ ਥਰੈਪੀ ਕੀ ਹੈ ਅਤੇ ਕਿਵੇਂ ਕੰਮ ਕਰਦੀ ਹੈ ਕਿਵੇਂ ਕਿਵੇਂ ਡਾਕਟਰ ਇਸ Therapy ਰਾਂਹੀ ਇਲਾਜ ਕਰਦੇ ਹਨ ... ਪੂਰਾ process ਸਮਝੋ ... ... Let me elaborate on the process of CAR T-cell therapy: 1. **T Cell Extraction:** - T cells, a type of white blood cell, are obtained from the patient's blood. - These T cells play a crucial role in the immune system by helping the body fight infections. 2. **Laboratory Modification:** - In a lab, scientists introduce a new gene into the T cells. - This gene encodes a special receptor known as a chimeric antigen receptor (CAR). - The CAR is designed to recognize and bind to specific proteins found on the surface of cancer cells. 3. **CAR T Cell Production:** - The modified T cells (now CAR T cells) are cultured and grown in large numbers. - These CAR T cells are armed with the ability to specifically target cancer cells due to the added CAR receptor. 4. **Infusion into the Patient:** - Once sufficient CAR T cells are produced, they are infused back into the patient's bloodstream. - These CAR T cells seek out cancer cells, bind to them, and initiate an immune response against the cancer. Overall, CAR T-cell therapy is a promising approach in cancer treatment, harnessing the patient's own immune system to target and destroy cancer cells. If you have any more questions or need further clarification, feel free to ask! 😊 ...

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https://lnkd.in/gUfz_DYt Native mass spec can be an excellent orthogonal validation technique to NMR for fragment campaigns.

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Enhancing RNA delivery by unraveling LNP fusion dynamics with advanced microscopy techniques. Something you may already know: LNPs' effectiveness largely hinges on the dynamic behaviors of ionizable cationic lipids, which adjust to pH variations through protonation/deprotonation processes. Now, if you are making LNPs, understanding these "rather complex" dynamics is essential for refining their performance and maximizing therapeutic delivery. However, there are still substantial gaps in our knowledge concerning how these particle formation mechanisms influence crucial aspects like drug loading and delivery. To bridge this knowledge gap, a new study has implemented an innovative approach using single-molecule convex lens-induced confinement (CLiC) microscopy alongside Förster resonance energy transfer (FRET) measurements (such the pair). This methodology allows us to observe LNP fusion dynamics in unprecedented detail, shedding light on how these particles interact under various conditions influenced by formulation parameters such as lipid concentration, buffer conditions, drug loading ratios, PEG-lipid concentrations, and the choice of ionizable lipids. The findings of this paper demonstrate a clear and consistent correlation between LNP fusion behaviors and the specific formulation parameters employed, confirming insights previously suggested by commonly-used techniques such as DLS and Cryo-TEM based assays. In summary, these advanced microscopy measurements not only provide a cost-effective way to characterize and screen potential drug candidates but also open new avenues for the targeted design and optimization of LNP-based delivery systems. By integrating these cutting-edge techniques, we can gain deeper insights into the fundamental processes governing LNP behaviors, ultimately leading to the development of more effective RNA delivery systems. Learn more about this here: https://lnkd.in/ehpjvmsb #Nanotechnology #RNAtherapeutics #VaccineDevelopment #Biotechnology #Microscopy #DrugDelivery #ScientificInnovation #HealthcareTechnology

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Researchers at UCLA & Georgia Institute of Technology unveil #RNAGPT, a multi-modal chat model that simplifies RNA discovery by leveraging extensive literature to streamline complex queries. 🧬 🎯 RNA-GPT integrates RNA sequence encoders with large language models to process user-uploaded sequences. According to the researchers, RNA-GPT effectively addresses complex RNA queries, helping researchers navigate the vast literature and unlock new insights with the power of multi-modal AI. Quick Read: https://lnkd.in/gAeEQJfZ #Bioinformatics #RNA #LLMs #ArtificialIntelligence #gpt #ScienceNews

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Researchers at UCLA & Georgia Institute of Technology unveil #RNAGPT, a multi-modal chat model that simplifies RNA discovery by leveraging extensive literature to streamline complex queries. 🧬 🎯 RNA-GPT integrates RNA sequence encoders with large language models to process user-uploaded sequences. According to the researchers, RNA-GPT effectively addresses complex RNA queries, helping researchers navigate the vast literature and unlock new insights with the power of multi-modal AI. Quick Read: https://lnkd.in/gAeEQJfZ #Bioinformatics #RNA #LLMs #ArtificialIntelligence #gpt #ScienceNews #biotechnology

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HEK293 Cells through Glycopeptide Mapping and Released Glycan Analysis Recombinant adeno-associated viral (AAV) vectors have emerged as prominent gene delivery vehicles for gene therapy. AAV capsid proteins determine tissue specificity and immunogenicity and play important roles in receptor binding, the escape of the virus from the endosome, and the transport of the viral DNA to the nuclei of target cells. Therefore, the comprehensive characterization of AAV capsid proteins is necessary for a better understanding of the vector assembly, stability, and transduction efficiency of AAV gene therapies. Glycosylation is one of the most common post-translational modifications (PTMs) and may affect the tissue tropism of AAV gene therapy. However, there are few studies on the characterization of the N- and O-glycosylation of AAV capsid proteins. In this study, we identified the N- and O-glycosylation sites and forms of AAV9 capsid proteins generated from HEK293 cells using liquid chromatography–tandem mass spectrometry (LC-MS)-based glycopeptide mapping and identified free N-glycans released from AAV9 capsid proteins by PNGase F using hydrophilic interaction. https://lnkd.in/e6z6TKHz

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A collaborative study involving Scripps Research has uncovered a critical vulnerability in Plasmodium falciparum, the parasite responsible for severe malaria. Researchers identified how broadly neutralizing antibodies (bnAbs) block a key site on the PfEMP1 protein, which is essential for the parasite’s survival in its host. This site, which cannot mutate without jeopardizing the parasite’s ability to infect, is a promising target for next-generation malaria vaccines. “Importantly, this study reveals how immunity can arise despite the parasite’s use of a highly mutated antigen to infect its host—a puzzle that has baffled scientists for many years,” says co-first author Sai Sundar Rajan Raghavan, PhD, from the Ward Lab at Scripps Research. Published in Nature Magazine, the study was conducted with collaborators from leading institutions, including the Københavns Universitet - University of Copenhagen; the UT Health San Antonio; the Fred Hutch Cancer Research Center in Seattle, EMBL Barcelona; the University of California, San Francisco; the Tanga Research Centre in Tanzania; and the Infectious Disease Research Centre in Uganda. Learn more at: https://ow.ly/MitX50Ue371

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bispecific Ab against PD-L1 and VEFG Ivonescimab, showed a 49% reduction in the risk of progression or death (stratified hazard ratio [HR], 0.51; 95% CI, 0.38 to 0.69; p<0.0001); in a phase 3 against Pembrlizumab in PD-L1-positive advanced NSCLC patients ( PFS 5.82 vs 11.14 months)

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Role of hMEX3B in Tumor Immune Evasion and RNA Regulation Immunotherapy, including immune checkpoint inhibitors and adoptive cell transfer, has made significant progress, but its effectiveness varies between patients due to genetic and epigenetic differences. A key factor in this variability is the human MEX3B (hMEX3B) protein, an RNA-binding protein involved in tumor immune evasion and regulation of HLA-A expression. hMEX3B contains two KH domains at its N-terminus and a RING domain at its C-terminus, which give it E3 ubiquitin ligase activity essential for RNA degradation. Despite its known biological importance, the specific RNA sequences recognized by hMEX3B and the underlying mechanisms of its function are not fully understood. Recently, in a study published in Communications Biology by Dr. Kanglong Yang et al., they screened the best hMEX3B binding sequence on HLA-A mRNA and found that two tandem KH domains had a 100-fold higher binding affinity to its target RNA than a single KH domain. Using nuclear magnetic resonance (NMR) and small-angle X-ray scattering (SAXS), they investigated the structural dynamics between the KH domain and its RNA substrate. Using molecular dynamics simulations, they developed a structural model of these interactions, identifying key regions on the KH domain (particularly two pairs of threonine and arginine) that significantly disrupt RNA recognition. These findings were validated both in vitro and in vivo. Their results suggest that hMEX3B plays a key role in regulating the degradation of its substrate mRNAs, affecting many biological processes, including immune responses to tumors. This study provides insight into the molecular mechanisms of hMEX3B and lays the foundation for studying the functions of other MEX3 family proteins. Understanding these mechanisms could help improve the effectiveness of immunotherapy by addressing the underlying molecular pathways involved in tumor immune evasion. Reference [1] Kanglong Yang et al., Communications Biology 2024 (https://lnkd.in/ePPjbVCE) #Immunotherapy #CancerResearch #MEX3B #RNARegulation #TumorEvasion #HLARegulation #BiomolecularResearch #CancerTreatment #RNADegradation #PrecisionMedicine #BiotechInnovation

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#Immunity | #Bcells | "Making the Transition" | Immature B-cell #Homing from the #BoneMarrow shapes Human #LymphoidTissue #Structure & #Function | Open Access 'Perspective' at Rockefeller University Press 👏 | #antibodies | Shortly after the emergence of newly formed human B cells from bone marrow as transitional cells, they diverge along two developmental pathways that can be distinguished by the level of IgM they express and migratory biases. Here*, the authors propose that differential tissue homing of immature B cell subsets contributes to human lymphoid tissue structure and function. *https://lnkd.in/eCQC9HVk Celentyx Ltd Professor Nicholas Barnes PhD, FBPhS Omar Qureshi Catherine Brady FIGURE | Recruitment of immature B cells to lymphoid tissues | B cells exit the bone marrow as T1 B cells, expressing high levels of IgM and CD10. Their transition to T2 cells is characterized by a reduction in CD10 expression and gain of CD21, with two distinct developmental pathways becoming apparent. T2 IgMhi cells selectively express α4β7 integrin, which mediates homing to GALT and spleen through binding to its cognate receptor MAdCAM-1. T2 IgMlo cells express relatively more L-selectin and are thus enriched in their potential to enter lymphoid tissues expressing its ligand PNAd, including lymph nodes and tonsils. We propose that T2 IgMhi and T2 IgMlo cells mature into MZB and naïve B cells, respectively. Higher receptor surface expression may enable IgMhi cells to respond to multivalent repeated subunit antigens that drive TI responses. IgMlo cells are instead adapted to T-dependent responses to single epitope antigens |

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Leverage our AI-powered platform for supercharged small molecule hit discovery. Watch now: https://lnkd.in/epSSRDte Learn how our Hits Discovery platform accelerates the discovery of potent, drug-like hit compounds with enhanced novelty and diversity with: - AI-driven Virtual Screening (VS) - High-Throughput Screening (HTS) - DNA-Encoded Library Screening (DEL)

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Two recent papers -- by Fojo and Olivier -- explained why Sotorasib, a KRAS G12C inhibitor, became a regulatory setback for NSCLC? They wrote: Sotorasib was approved on the basis of a single-arm, uncontrolled phase 1–2 study (CodeBreaK 100) that reported an objective response rate (ORR) of 37% and a 11·1 month median duration of response. By contrast, other molecular therapies for NSCLC have higher ORRs; osimertinib targets EGFR-mutated NSCLC and has an ORR of 90% and alectinib targets ALK-mutated NSCLC and has reported an ORR of 82·9%.

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