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ADCC/CDC Enhancement in Therapeutic Antibody Development
Therapeutic antibodies, engineered through biotechnology, represent a specialized class of antibodies used in disease treatment. These antibodies are designed to target specific disease markers, such as malignant tumors, autoimmune disorders, and infectious diseases. Compared to traditional antibody therapies, therapeutic antibodies offer higher specificity and fewer side effects. In the realm of immunotherapy, antibody-dependent cell-mediated cytotoxicity (ADCC) stands out as a highly effective anti-tumor mechanism. ADCC enhancement refers to the bolstering of immune cells' ability to attack malignant cells, thereby enhancing the efficacy of immunotherapy. ADCC enhancement technology finds significant applications in the field of therapeutic antibodies, encompassing techniques like fucosylation engineering, Fc protein-engineering, cross-isotype engineering, and glyco- and Fc protein dual engineering. Furthermore, antibody-dependent cell phagocytosis (ADCP) plays a pivotal role in the action of therapeutic antibodies. The ADCP assay serves as an experimental method for studying antibody-dependent cell phagocytosis. This research investigates whether antibodies assist immune cells, such as macrophages, in recognizing, engulfing, and digesting labeled target cells or pathogens. Through the ADCP assay, researchers can assess whether therapeutic antibodies activate immune cells to attack and eliminate tumor cells, instilling renewed optimism in cancer treatment. CDC enhancement, a classical approach to fortifying the immune system, amplifies the cytotoxicity of antibodies. Immunotherapy often hinges on antibody action, and CDC enhancement accentuates the activation of the complement system by antibodies, inducing cell toxicity and ultimately eradicating target cells. In CDC enhancement, antibodies (typically therapeutic monoclonal antibodies) bind to antigens on the surface of target cells, triggering the activation of the C1q molecule in the complement system. C1q further instigates the complement cascade reaction in the immune system, culminating in the formation of the membrane attack complex (MAC). This process ruptures target cell membranes and leads to cell lysis, achieving cytotoxic effects on the target cells. Researchers assess the binding capacity of therapeutic antibodies with C1q through the C1q binding assay, determining the antibody's effectiveness in the immune response. Researchers have surmounted numerous challenges in disease treatment through advanced techniques such as the C1q binding Assay and ADCP assay. In cancer treatment, scientists have successfully developed a series of antibodies targeting specific antigens. These drugs activate immune cells, propelling them to engulf and annihilate cancer cells. The successful application of this immunotherapy brings renewed hope to tumor treatment. In the domain of autoimmune disease treatment, researchers are leveraging antibodies to target diseases resulting from immune system overactivation. Through meticulous C1q binding assay studies, scientists can pinpoint the most suitable antibodies for treatment, precisely modulating the immune system's activity to achieve therapeutic goals. Moreover, in the realm of treating viral and bacterial infections, the utilization of the ADCP assay is on the rise. Researchers have formulated a series of antibodies targeting pathogens, effectively eliminating infection sources by stimulating immune cells to engulf these pathogens. Consequently, this approach has significantly heightened the success rate of infectious disease treatments. With the continuous evolution of single-cell technologies and CRISPR gene editing techniques, researchers can delve deeper into cell death mechanisms, antibody structures, and immune cell functions. This progress will further accelerate research on ADCC enhancement, offering more precise and efficient means for disease treatment.
Cell Therapy: Surface Modification Technology Based on Cell Membrane
The cell membrane acts not only as a physical barrier but also as a functional organelle that regulates communication between cells and their environment. Functionalizing the cell membrane using synthetic molecules or nanostructures has the potential to enhance cellular functions beyond those achieved through natural evolution. Cell therapy represents a groundbreaking approach in treating major challenging diseases, including tissue injuries, degenerative diseases, and congenital metabolic disorders. The primary focus of biomedical research has always been on regulating cellular functions to maximize the efficiency of cell therapy. Given that the cell surface plays a critical role in cellular physiology and pathology by controlling recognition and communication between cells and their environment, functionalizing the cell surface emerges as an effective method for regulating cellular functions. We have developed a range of cell surface modification techniques based on molecular self-assembly approaches, wherein exogenous biomolecules and biomaterials are constructed on the cell surface through molecular engineering to regulate cell function and enhance the efficacy of cell therapy. This non-genetic engineering-based modification of the cell surface can functionalize cells within hours, significantly reducing manufacturing costs and processes without genetically modifying the cells, thereby making transient manipulation of cell functions feasible while avoiding potential safety risks. The highly specific biotin-avidin interaction exhibits remarkable resistance to harsh denaturing conditions, including heat, pH fluctuations, and organic solvents. Consequently, biotinylation holds immense promise in cell surface engineering. Cell surface-based biotinylation modification, leveraging the strong affinity between biotin molecules and avidin, enables the specific introduction of biotin on the cell surface, thereby functionalizing the cell through biotin-avidin binding. This technology typically involves the following steps: Introduction of biotin linker: Initially, molecules containing biotin linker groups must be introduced onto the cell surface. This can be accomplished through various methods, such as employing compounds containing biotin or utilizing biotin ligase to catalyze the covalent binding of biotin to cell surface molecules. Covalent binding of biotin linker with cell surface molecules: The biotin linker forms covalent bonds with molecules on the cell surface, thereby introducing biotin onto the cell membrane. This binding is typically highly specific, enabling the selective modification of specific cell surface structures. Interaction between biotin and avidin: Once the cell surface is labeled with biotin, the high -affinity interaction between biotin and avidin is utilized to functionalize the cell. Avidin is usually associated with fluorescent labels, polymers, or other molecular tags, which, upon specific binding with biotin, are introduced onto the cell surface, achieving functional modification of the cell. Functional application: Following the labeling of the cell surface with biotin and its binding to avidin, various functional modifications of the cell can be achieved. For instance, fluorescent labels can be utilized for cell imaging, drug carriers can be attached to the cell surface for drug delivery, or other functional molecules can be employed to regulate cell signaling, among other applications. Utilizing cell membrane coating technology to enhance the efficacy of drugs involves introducing additional cell membrane functions to increase their specificity. Although cell membrane-coated nanoparticles (CM-NPs) can achieve prolonged circulation, adding targeting ligands can enhance their localization to specific targets, such as tumors. This cell membrane-based ligand modification technology offers a simpler and more effective approach by combining natural cell membranes with different ligands for biological tasks. This strategy involves stabilizing functional ligand molecules on the extracellular domains of cell membrane proteins using cell-impermeable linkers. The crux of this method lies in coupling the ligand with cell membrane proteins, thereby achieving functional modification of the cell membrane. This cell membrane-based surface engineering technology offers drug delivery systems with enhanced specificity and targeting, particularly in fields like tumor therapy, with extensive application prospects.
Bispecific Antibodies: A Rising Force in Revolutionary Cancer Treatment
Immunotherapy stands out as the most promising systemic approach to cancer treatment compared to conventional methods. Monoclonal antibodies, known for their ability to precisely target molecules, have emerged as a vital and effective modality in cancer therapy. However, the intricacies of tumor development often limit the effectiveness of monoclonal antibodies targeting a single point. The introduction of bispecific antibodies (bsAbs), capable of targeting multiple sites simultaneously, has transformed the landscape of tumor immunotherapy. What is a bispecific antibody? Over the last few decades, there has been a notable shift from developing and modifying basic antibodies (Abs) to more intricate Ab derivatives, with a special focus on bsAbs of varied shapes and sizes. BsAb technology holds tremendous promise in clinical applications, garnering researchers' attention and evolving into diverse forms, establishing a robust foundation for cancer immunotherapy centered around bsAbs. Presently, a multitude of preclinical and clinical trials are underway, marking the era of bispecific antibodies in tumor immunotherapy. As of December 2021, the United States Food and Drug Administration (FDA) has granted approval for three types of bsAbs for clinical cancer treatment. Due to their capability to simultaneously target two epitopes on tumor cells or within the tumor microenvironment (TME), bsAbs have become a pivotal and promising element of the next generation of therapeutic antibodies. The majority of bsAbs in current development are crafted as T-cell engagers, forging close connections between immune cells, particularly cytotoxic T cells, and tumor cells to create an artificial immune contact. This ultimately leads to selective attacks and lysis of targeted tumor cells. Bispecific T-cell engagers, as a groundbreaking cancer immunotherapy strategy, have exhibited encouraging results in clinical trials, particularly in hematologic malignancies. To date, only one bispecific T-cell engager, blinatumomab, has received approval from the FDA and the European Medicines Agency for treating relapsed or refractory B-cell precursor acute lymphoblastic leukemia (B-ALL) and minimal residual disease (MRD)-positive B-ALL. Additionally, numerous other bispecific T-cell engagers are undergoing clinical trials, targeting various tumor types, including hematologic malignancies and solid tumors. Classified by their functional mechanisms, bsAbs, besides cell-cell engagers, can be further divided into those binding two epitopes on the same antigen, dual-functional modulators, and bsAbs in cell therapy. One innovative form includes those with an antigen-binding Fc fragment (Fcab), incorporating a homodimeric Fc region with antigen-binding sites. This distinctive combination enables Fcabs to simultaneously leverage the functions mediated by the Fc domain and antigen-binding capabilities. Significantly, Fcabs are one-third smaller than full-length antibodies, facilitating superior tissue penetration, particularly advantageous in treating solid tumors. Moreover, Fcabs serve as a robust foundation for creating antibody-drug conjugates (ADCs), ensuring precise drug delivery by linking cytotoxic drugs specifically to Fcab. While most bsAbs in clinical trials presently target hematologic malignancies, exploring bsAbs targeting solid tumors is essential due to their inevitable adverse effects on normal tissues. Factors like immune-tolerant cancer stroma, angiogenic disorders, and insufficient penetration of bsAb drugs contribute to the complexity of this exploration. As a result, there is enthusiastic interest in ongoing research on bsAbs in solid tumors. In conclusion, the outcomes of bsAb research underscore the promising prospects of these molecules in innovative drug design and subsequent clinical applications in cancer treatment.
Deciphering the Importance of Single-Cell Sequencing
The term "single cell" refers to an individual cell, isolated and examined on its own. Analysis conducted specifically on individual cells is collectively referred to as single-cell sequencing analysis, while sequencing performed on these isolated cells is termed single-cell sequencing. Sequencing multiple cells or a group of cells falls outside the realm of single-cell sequencing. For instance, common genetic sequencing practices, often performed for public interest, entail extracting specific DNA fragments after minimal blood processing. However, it remains uncertain whether the extracted DNA originates from a particular white blood cell, another white blood cell, or free DNA circulating in the bloodstream. Similarly, in conventional tumor studies, sequencing is typically conducted on numerous tumor cells isolated from tumor tissue. Single-cell sequencing for oncology represents a specialized form of sequencing; currently, the majority of sequencing efforts do not operate at the single-cell level. To grasp the technical aspects of single-cell sequencing and analyze its advantages, it's crucial to understand the precise meanings of terms such as "single-cell sequencing" and "high-throughput technology." We need to discern what these terms entail when prefixed with "single cell" or "high-throughput." The fundamental significance of single-cell sequencing lies in cellular heterogeneity. This implies that individual cells exhibit variability, even among cells from the same location, potentially resulting in differences in gene expression and other attributes. Studying cell populations only provides averaged outcomes, masking cellular heterogeneity. Two specific examples illustrate this: Firstly, cell classification. Historically, cell classification relied on characteristics like spatial position and morphology, which is a relatively crude method. Conducting single-cell RNA or DNA sequencing enables a more nuanced and rigorous cell classification, particularly beneficial for complex tissues, facilitating a deeper understanding of cellular functions. Secondly, studies related to tumors. A widely accepted hypothesis regarding tumor metastasis posits that certain cells from a tumor may detach, enter the bloodstream, and become circulating tumor cells (CTCs). Some CTCs may travel to an organ via the bloodstream, invade blood vessels, infiltrate the organ, adhere, proliferate, and form new tumors. Determining which cells from the original tumor become CTCs, which CTCs can survive in the bloodstream, and complete tumor metastasis requires single-cell level sequencing and other related research endeavors. In conclusion, the advent of single-cell sequencing has opened new vistas in our understanding of cellular biology, particularly in unraveling the complexities of cellular heterogeneity. By delving into the intricacies of individual cells, we can uncover insights that were previously obscured by population-level analyses. This approach holds immense promise in various fields, from advancing our knowledge of basic cellular functions to revolutionizing our understanding of diseases like cancer. As we continue to refine and expand single-cell sequencing technologies, we can anticipate even greater breakthroughs on the horizon, unlocking the full potential of this powerful tool in biological research and clinical practice.
Breakthrough mRNA Research Garners 2023 Nobel Prize in Physiology or Medicine
On October 2nd, the Nobel Assembly unveiled the recipients of the 2023 Nobel Prize in Physiology or Medicine: scientists Katalin Karikó and Drew Weissman. Their pioneering research in messenger ribonucleic acid (mRNA) has reshaped vaccine development, notably amid the COVID-19 pandemic. Their work has not only saved countless lives but has also alleviated the severity of cases, relieving pressure on healthcare systems and facilitating the global reopening of societies. To date, mRNA vaccines, administered over 13 billion times worldwide, have played a pivotal role in fighting the pandemic. Scientists have delved into mRNA's potential for vaccine development since the 1990s. The laureates' work "revolutionized our comprehension of mRNA's interaction with the immune system," crucial in the swift creation of mRNA vaccines for SARS-CoV-2 during the ongoing global health crisis. These vaccines deliver the spike protein mRNA sequence into cells using lipid nanoparticles (LNPs) as carriers. This innovative method triggers protein production, activating immune cells and eliciting responses like the creation of neutralizing antibodies and antigen-specific T cells. mRNA-based SARS-CoV-2 vaccines boast rapid production and cost-effectiveness. By amplifying antigens through mRNA synthesis, high concentrations of neutralizing antibodies are achieved, enhancing vaccine efficacy. In contrast, producing vaccines based on whole viruses or viral proteins necessitates extensive cell cultures, complicating rapid pandemic vaccine production. SARS-CoV-2 vaccine candidates underwent testing in animal models like ACE2 humanized mice, ferrets, and rhesus macaques. Exogenous mRNA corresponding to viral gene fragments enables host cells to produce viral proteins, stimulating immune responses and serving as vaccine candidates. Yet, extracellular mRNA production suffers from instability and inefficient delivery. The laureates' research showcased that modifying extracellular mRNA's nucleotide bases could make the host "recognize" exogenous mRNA as self-mRNA. This modification reduces inflammatory reactions and boosts protein production after delivery, removing key hurdles in mRNA's clinical application. This breakthrough paves the way for agile mRNA vaccine development for infectious diseases and holds potential for delivering therapeutic proteins and treating specific cancer types However, mRNA is inherently unstable and prone to enzymatic degradation within the body. Another challenge lies in the potential for mRNA to trigger intense inflammatory responses, potentially harming cells and tissues. Despite skepticism and rejection, Karikó and Weissman persevered. In 2005, they published a groundbreaking paper addressing these challenges. By modifying mRNA's building blocks, nucleotides, they enhanced stability and reduced immunogenicity. Additionally, they devised a method employing lipid nanoparticles to deliver mRNA into cells, safeguarding and transporting mRNA within minuscule lipid bubbles. Karikó and Weissman's work stands as a groundbreaking transformation in anti-SARS-CoV-2 candidates and public health, illustrating the potency of curiosity-driven science and resilience. Their achievements inspire researchers and innovators worldwide to explore mRNA technology's potential in enhancing human health and well-being.
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