Monoclonal Antibody Technology Research Articles

Clinical Diagnostic Immunology in A Clinical Microbiology Laboratory

This review unravels the novel approaches, innovations and applications of immunology in the diagnosis of infections in a clinical microbiology laboratory. Many assays or tests in immunology are used in the diagnosis of infections. The availability of monoclonal antibodies has brought rapid progress to this area and they are used , for example , to identify specific microorganisms in culture or in tissues and fluids from infected patients. Examples of common diagnostic applications of antibody- antigen reactions discussed here includes agglutination tests, complement tests, complement fixation tests, ELISA, cytotoxic tests, radioimmunoassay tests, haemagglutination inhibition tests, hypersensitivity tests, phagocytic cell activity test, migration inhibition test, lymphocyte transformation test , lymphocyte fixation test , skin tests, delayed hypersensitivity tests and many others. The development of monoclonal antibody technology and the subsequent availability of unlimited quantities of pure antibody has improved the selectivity of these assays. The development of simple and highly sensitive methods for detection of nucleic acids is being applied to diagnosis of infection using nucleic acid hybridization and this provides a valuable additional aid to immunoassays in the clinical microbiology diagnostic laboratory.

Establishment of colloidal gold immunochromatography strip for rapid detection of Karen mikimotoi and its application.

In China, harmful algal blooms (HABs) are one of the most prominent ecological disasters in the coastal areas. Among the harmful algae species that cause HABs, Karen mikimotoi is a kind of algae that appear frequently. It can secrete hemolytic toxins and fish toxins such as glycolipids and unsaturated fatty, posing a significant threat to marine life. In order to establish a fast and effective detection technology for Karen mikimotoi that can be promoted and applied on site, we have developed a test strip which is based on monoclonal antibody technology and the colloidal gold immune-chromatography assay (GICA). The experimental results show that this test strip can detect different growth stages including growth, and stable and recession period of Karen mikimotoi. The detection results can be displayed within 3-15min. It had high sensitivity and specificity, with a detection limit of 754 cells/mL. A colorimetric card was made to further determine the concentration of algae detected. What is more, we had developed a method that can be used for on-site enrichment of algae cells using a syringe to detect lower concentrations of Karen mikimotoi, with a minimum detection concentration of 100 cells/mL. Also the test strip was used for on-site testing along the coast of China. This test strip not only serves as a warning for the outbreak of red tide, but also provides a new approach for the development of rapid detection technology for red tide algae.

Antibodies as Models and Tools to Decipher Candida albicans Pathogenic Development: Review about a Unique Monoclonal Antibody Reacting with Immunomodulatory Adhesins.

Candidiasis, caused mainly by Candida albicans, a natural commensal of the human digestive tract and vagina, is the most common opportunistic fungal infection at the mucosal and systemic levels. Its high morbi-mortality rates have led to considerable research to identify the molecular mechanisms associated with the switch to pathogenic development and to diagnose this process as accurately as possible. Since the 1980s, the advent of monoclonal antibody (mAb) technology has led to significant progress in both interrelated fields. This linear review, intended to be didactic, was prompted by considering how, over several decades, a single mAb designated 5B2 contributed to the elucidation of the molecular mechanisms of pathogenesis based on β-1,2-linked oligomannoside expression in Candida species. These contributions starting from the structural identification of the minimal epitope as a di-mannoside from the β-1,2 series consisted then in the demonstration that it was shared by a large number of cell wall proteins differently anchored in the cell wall and the discovery of a cell wall glycoplipid shed by the yeast in contact of host cells, the phospholipomannan. Cytological analysis revealed an overall highly complex epitope expression at the cell surface concerning all growth phases and a patchy distribution resulting from the merging of cytoplasmic vesicles to plasmalema and further secretion through cell wall channels. On the host side, the mAb 5B2 led to identification of Galectin-3 as the human receptor dedicated to β-mannosides and signal transduction pathways leading to cytokine secretion directing host immune responses. Clinical applications concerned in vivo imaging of Candida infectious foci, direct examination of clinical samples and detection of circulating serum antigens that complement the Platelia Ag test for an increased sensitivity of diagnosis. Finally, the most interesting character of mAb 5B2 is probably its ability to reveal C. albicans pathogenic behaviour in reacting specifically with vaginal secretions from women infected versus colonized by this species as well as to display higher reactivity with strains isolated in pathogenic circumstances or even linked to an unfavourable prognosis for systemic candidiasis. Together with a detailed referenced description of these studies, the review provides a complementary reading frame by listing the wide range of technologies involving mAb 5B2 over time, evidencing a practical robustness and versatility unique so far in the Candida field. Finally, the basic and clinical perspectives opened up by these studies are briefly discussed with regard to prospects for future applications of mAb 5B2 in current research challenges.

Selection of phage-displayed antibodies with high affinity and specificity by electrophoresis in microfluidic devices.

A method development aimed for high-throughput and automated antibody screening holds great potential for areas ranging from fundamental molecular interactions to the discovery of novel disease markers, therapeutic targets and monoclonal antibody engineering. Surface display techniques enable efficient manipulation of large molecular libraries in small volumes. Specifically, phage display appeared as a powerful technology for selecting peptides and proteins with enhanced, target-specific binding affinities. Here, we present a phage-selection microfluidic device wherein electrophoresis was performed under two orthogonal electric fields through an agarose gel functionalized with the respective antigen. This microdevice was capable of screening and sorting in a single round high affinity phage-displayed antibodies against virus glycoproteins, including Human Immunodeficiency Virus-1 (HIV-1) glycoprotein (gp120) or the Ebola Virus glycoprotein (EBOV-GP). Phages were differentially and laterally swept depending on their antigen affinity; the high affinity phages were recovered at channels proximal to the application site, whereas low affinity phages migrated distal after electrophoresis. These experiments proved that the microfluidic device specifically designed for phage-selection is rapid, sensitive and effective. Therefore, this is an efficient and cost-effective method that allowed highly controlled assay conditions for isolating and sorting high affinity ligands displayed in phages. This article is protected by copyright. All rights reserved.

Monoclonal Antibodies in Cancer Immunotherapy

The technology of monoclonal antibodies (MABs) was originated from the ideas that how antibodies of the B cells bind to the specific antigens to protect the body against foreign invasion. Nowadays, MABs drugs have been approved for more than 30 targets, they have been considered as one of the first-line immunotherapies which specifically target certain cancers. Due to their exquisite target selectivity and less toxicity, MABs have become the new mainstay of pharmaceutical industry. This review summarizes the history of discovery, basic structure, and in vivo production of the MABs. Meanwhile, this review introduces the basic therapeutic applications, mechanism and working process of MABs, MABs and its targeted cancers, issues with current MABs technology and corresponding methods of improvements are also discussed. Overall, MABs do have broad prospect in the future since its effectiveness of treating cancers with poor prognosis and wide therapeutic applications.

Antibodies to Combat Fungal Infections: Development Strategies and Progress

The finding that some mAbs are antifungal suggests that antibody immunity may play a key role in the defense of the host against mycotic infections. The discovery of antibodies that guard against fungi is a significant advancement because it gives rise to the possibility of developing vaccinations that trigger protective antibody immunity. These vaccines might work by inducing antibody opsonins that improve the function of non-specific (such as neutrophils, macrophages, and NK cells) and specific (such as lymphocyte) cell-mediated immunity and stop or aid in eradicating fungus infections. The ability of antibodies to defend against fungi has been demonstrated by using monoclonal antibody technology to reconsider the function of antibody immunity. The next step is to develop vaccines that induce protective antibody immunity and to comprehend the mechanisms through which antibodies mediate protective effects against fungus.

Progress and perspectives of rabbit monoclonal antibodies

Since the advent of murine hybridomas, the emergence of a variety of monoclonal antibody (mAb) technologies has enabled the wide applications of murine monoclonal antibodies in medicine, life science, agronomy, and food science. Compared with murine monoclonal antibodies, rabbit monoclonal antibodies (RabmAbs) exhibit higher affinity, presenting with increased detection sensitivity and greater specificity for the particular structure of epitopes. This paper reviews the history, preparation techniques, advantages and disadvantages, current applications, and future perspectives for RabmAbs.

Isolation and characterization of full-length genes encoding the anti-human CD45 antibody from the hybridoma cell line 16E8-F2

Though the hybridoma technology has been widely applied in the production of monoclonal antibodies, it has existed some disadvantages including low yield and genetic instability. Therefore, an alternative approach should be taken into account. Recently, recombinant monoclonal antibody technology has emerged as the best choice to cure hybridoma related drawbacks. However, recombinant antibodies require known genes for their generation. The purpose of this study is to collect the full-length genes encoding the anti-human CD45 antibody derived from the hybridoma cell line 16E8-F2. In this research, we designed specific primer pairs to amplify the light and heavy chain genes of the antibody through the PCR method. Afterward, the genes were separately cloned into a cloning vector called pJET1.2/blunt. The generated recombinant pJET1.2 vectors will serve as the main material source to manufacture the recombinant monoclonal antibody recognizing human CD45 protein tomorrow.

Monoclonal Antibody Engineering and Design to Modulate FcRn Activities: A Comprehensive Review.

Understanding the biological mechanisms underlying the pH-dependent nature of FcRn binding, as well as the various factors influencing the affinity to FcRn, was concurrent with the arrival of the first recombinant IgG monoclonal antibodies (mAbs) and IgG Fc-fusion proteins in clinical practice. IgG Fc–FcRn became a central subject of interest for the development of these drugs for the comfort of patients and good clinical responses. In this review, we describe (i) mAb mutations close to and outside the FcRn binding site, increasing the affinity for FcRn at acidic pH and leading to enhanced mAb half-life and biodistribution, and (ii) mAb mutations increasing the affinity for FcRn at acidic and neutral pH, blocking FcRn binding and resulting, in vivo, in endogenous IgG degradation. Mutations modifying FcRn binding are discussed in association with pH-dependent modulation of antigen binding and (iii) anti-FcRn mAbs, two of the latest innovations in anti-FcRn mAbs leading to endogenous IgG depletion. We discuss the pharmacological effects, the biological consequences, and advantages of targeting IgG–FcRn interactions and their application in human therapeutics.

Retrospective analysis of the preparation and application of immunotherapy in cancer treatment (Review).

Monoclonal antibody technology plays a vital role in biomedical and immunotherapy, which greatly promotes the study of the structure and function of genes and proteins. To date, monoclonal antibodies have gone through four stages: murine monoclonal antibody, chimeric monoclonal antibody, humanised monoclonal antibody and fully human monoclonal antibody; thousands of monoclonal antibodies have been used in the fields of biology and medicine, playing a special role in the pathogenesis, diagnosis and treatment of disease. In this review, we compare the advantages and disadvantages of hybridoma technology, phage display technology, ribosome display technology, transgenic mouse technology, single B cell monoclonal antibody generation technologies, and forecast the promising applications of these technologies in clinical medicine, disease diagnosis and tumour treatment.

“(Green)Biotechnology & Food Security”

This world and its nations are moving towards and working towards creation and formulation of “BioEconomy”, an economy which revolves around bioscience and biotechnology products and consumers. Since centuries and more importantly since last few decades in this modern and scientific era, the people of this world, its corporations, research laboratories as well as governments of different countries are working towards creation of Biotech products, drugs, Genetically Modified Foods, selectively breeding’s, Genetically modified grains and seeds to help evolve and create modified species, food and grains which could be cultivated in the toughest terrain to eco-friendly and sustainable methods of agro production and cultivations with a aim towards creating green biotechnology and finding solution of food scarcity and how the global crisis of food security and its supply and demand can be addressed. Moreover, in reality of this 21st century, people of this century have been successful in creation of Genetically Modified grains, plants, fruits, foods, and other agro products. And now we are moving towards commercialization of these genetically modified or agro products created by using biotechnology. However, before going further, the researcher would like to clarify that the scope of the research article is limited to biotechnology and how it can help address the problem of food security or food insecurity. It will try and explore the concept of green biotechnology and how it can help reduced food scarcity and the needs can be met with of food production and supply within the countries such as India, Africa etc. where poverty, unemployment and food scarcity or hunger are issues which exist at large scale. The aim of the research article is to explore the biotechnology as a solution for providing food security and to explore its various aspects in brief. Biotechnology and Food Security: Feeding the demographics in the 21st century, it has been estimated that to feed people basic essential food in the year 2025 average yield must increase by 80% than of the average yield in the year 1990. Which is a ginormous task, and a question that whether can this be achieved. Thus feeding food in the 21st century along with the current rate of poverty, unemployment and other problems is a difficult task to achieve and these task are the most essential for a peaceful survival of human race, which have to be addressed. Thus here agriculture biotechnology, genetic revolution comes into the picture. Feeding people for the next 25 years require inclusion or inclusion of genetic technologies, development and research on formulation and creation of green biotechnology. Here it is pertinent to note that biotechnology has to play the most pertinent role, because green biotechnology or biotechnology has an ability to directly modify plant, animals and agricultural process to resolve the current needs and issues of food security. The current work in the field of agro biotechnology or green biotechnology is development of seeds and plants which create more yield, needless fertilizers and water and also can sustain the change in the weather and environment. To help better understand, here in under the researcher has to tried to incorporate certain methods, products and techniques of green biotechnology or agro biotechnology which will help resolve the issue of food security and will be able to feed the 21st century. These methods, techniques and products are: “enzyme technology and monoclonal antibody technology; bio fertilizers, biocontrol agents and diagnostics; digestion metabolism; drug and vaccine development; selection and breeding of tissue culture, genetic markers, genetic modification, mutation induction, somatic hybridization and cybridization; mass propagation; detection of hormones level; embryo technology; germ plasm conservation” etc.

Preparation and application of yellow fever virus NS1 protein-specific monoclonal antibodies.

Yellow fever is an acute infectious disease that is common in Africa and South America and causes thousands of deaths annually. However, there are very few studies on yellow fever virus (YFV) antigen detection kits. As a detection target, the nonstructural protein 1 (NS1) has been successfully used in the early diagnosis of dengue virus (a member of the Flaviviridae family) infection. In this study, we used monoclonal antibody technology to prepare anti-YFV NS1 monoclonal antibodies (MAbs) and identified their immunological properties. Next, we used two mouse MAbs that can recognize different epitopes of YFV NS1 as capture and detection antibodies to establish a YFV NS1 antigen-capture enzyme-linked immunosorbent assay (ELISA). The antigen-capture ELISA displayed exclusive specificity to YFV without cross-reaction with other related members of the flavivirus family, including the dengue virus, West Nile virus, Japanese encephalitis virus. Additionally, the detection sensitivity towards the YFV culture supernatant was 103 TCID50/mL and the detection positivity rate was 95% compared with reverse transcription-polymerase chain reaction. In conclusion, this newly developed NS1 antigen-capture ELISA with high sensitivity and specificity could be used as an efficient method for the early diagnosis of YFV infection in animals or humans.

Lipid-Lowering Biotechnological Drugs: from Monoclonal Antibodies to Antisense Therapies-a Clinical Perspective.

While low density lipoprotein cholesterol (LDL-C) remains a key contributor of atherosclerotic cardiovascular disease (ASCVD), additional risk factors identified through epidemiological and genetic studies have ushered in a fertile era of drug discovery in lipid-lowering therapy. Unlike contemporary small molecule medications, many of the novel agents are biologics utilizing monoclonal antibody (mAb) or RNA interference (RNAi) technologies. This report aims to review the evidence to date, focusing on completed and ongoing clinical trials and how these new agents will impact clinical practice. We review data from pertinent studies on lipid-lowering biologics in clinical use or have translated to human studies and are undergoing clinical trials. Several targets affecting lipid metabolism have been identified to be causally associated with ASCVD including proprotein convertase subtilisin/kexin type 9 (PCSK9), angiopoietin-like protein 3 (ANGPTL3), apolipoprotein C3 (APOC3), and lipoprotein (a) (Lp[a]). Biotechnological modalities that have been developed for these targets include mAb, small interfering RNA (siRNA), and anti-sense oligonucleotide (ASO) agents. Agents such as alirocumab and evolocumab have shown efficacy in risk reduction of ASCVD in cardiovascular outcome trials and have been incorporated into evidence-based practice guidelines. Other agents included in this review are in various stages of clinical trials and have shown significant efficacy in the reduction of lipid parameters. The development of new biologics targeting lipid risk factors will provide clinicians additional tools to reduce the risk for ASCVD. Important factors to consider will be cost-effectiveness and improving methods to personalize treatments to risk factors.

Monoclonal Antibody-Based Therapies for Myasthenia Gravis.

Myasthenia gravis (MG) is an autoimmune, neuromuscular disorder that produces disabling weakness through a compromise of neuromuscular transmission. The disease fulfills strict criteria of an antibody-mediated disease. Close to 90% of patients have antibodies directed towards the nicotinic acetylcholine receptor (AChR) on the post-synaptic surface of skeletal muscle and another 5% to the muscle-specific kinase, which is involved in concentrating the AChR to the muscle surface of the neuromuscular junction. Conventional treatments of intravenous immunoglobulin and plasma exchange reduce autoantibody levels to produce their therapeutic effect, while prednisone and immunosuppressives do so by moderating autoantibody production. None of these treatments were specifically developed for MG and have a range of adverse effects. The extensive advances in monoclonal antibody technology allowing specific modulation of biological pathways has led to a tremendous increase in the potential treatment options. For MG, monoclonal antibody therapeutics target the effector mechanism of complement inhibition and the reduction of antibody levels by FcRn inhibition. Antibodies directed against CD20 and signaling pathways, which support lymphocyte activity, have been used to reduce autoantibody production. Thus far, only eculizumab, an antibody against C5, has reached the clinic. We review the present status of monoclonal antibody-based treatments for MG that have entered human testing and offer the promise to transform treatment of MG.

Development of enzyme linked immunosorbent assay of aflatoxin of Chinese herbal medicines

The enzyme linked immunosorbent assay of aflatoxin has been adopted in Chinese Pharmacopoeia(2020 edition). Based on high-throughput screening of monoclonal antibodies technology, monoclonal antibodies that can specifically recognize the aflatoxin B_1 and the total amount of aflatoxin B_1, B_2, G_1, and G_2 in Chinese herbal medicines were prepared. By optimizing the concentration of coating antibody, enzyme-labeled antigen, and the reaction system of enzyme-linked immunosorbent assay, the enzyme linked immunosorbent assay(ELISA) were developed for detection of aflatoxins in Chinese herbal medicines, decoction pieces, and preparation of Chinese medicine. In this method, the recovery test of actual samples is 60%-120%, and the relative standard deviation is less than 15%. In addition, in view of the complicated and expensive pretreatment methods for the determination of aflatoxin in Chinese herbal medicine, we developed a highly efficient pretreatment method of liquid-liquid extraction of aflatoxin in Chinese herbal medicine without immunoaffinity column. As an effective method for the detection of aflatoxin, the ELISA can effectively reduce the aflatoxins testing cost of traditional Chinese medicine, and promote the detection ability at earlier stages of production, and strengthen the quality supervision of traditional Chinese medicine.

Options for COVID-19 therapeutics: Aerosolized inhalation antibody-conjugated Nano particles.

The world is currently faced with a very serious crisis to deal with the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or Covid-19) pandemic which started in Wuhan, China in December 2019 and has since spread throughout the world, wreaking havoc in many countries. Several efforts are being made to control the spread of the disease around the world and to find a cure or vaccine. As researchers frantically endeavor to identify remedies for covid19, there is the need to identify therapies that offer the quickest, safest actions and remedies that are relatively cheap. We propose the use of aerosolized inhalation antibody conjugated nanoparticles for the treatment of covid-19. It is hypothesized in this proposal that the conjugation of nanoparticles with antibodies and delivering the antibody-nanoparticle conjugate as an aerosol via the respiratory tract would provide the quickest and possibly more efficient and relatively cheap remedy against covid-19. The advantage of the inhalation route for delivering antibody conjugated nanoparticles is that since the medication is delivered directly to the affected site, higher doses will be delivered to the site with reduced systemic toxicity and reduced adverse effects on gaseous exchange. Our hypothesis is based on the current knowledge and observations in the areas of monoclonal antibody technology, advances in nanotechnology and Nano medicine as well as advances in inhalation therapeutics.

Abstract A08: Development of FGFR4-specific chimeric antibody receptor (CAR) T cell and bispecific T cell engager (BiTE) for rhabdomyosarcoma (RMS) immunotherapy

Abstract Background: Despite decades of multimodule therapies, RMS remains incurable once it has metastasized; thus, new therapeutic strategies are warranted. FGFR4 is a developmentally regulated cell surface receptor tyrosine kinase, overexpressed in virtually all, mutationally activated in about 7.5% of RMS, and directly activated by PAX3-FOXO1 fusion protein, which makes it a tractable target for immunotherapy. Material and Methods: Using monoclonal antibody technologies and a yeast display B-cell library, we generated 15 human or mouse binders specific to human FGFR4 and engineered into human scFvFc. All binders were successfully produced in vitro, and we further characterized them using FACS and ELISA for their specificity. Octet was used to measure the binding affinity against human FGFR4. For those lead hits, they were made into different formats of therapeutic including CAR and BiTE. We then performed in vitro killing assays and/or in vivo xenograft model to determine the efficacy of those therapeutics in killing RMS cells. Results: m410 and m412 were two lead hits and scFvFcs of these two binders were successfully produced in vitro and showed FGFR4 specificity with a binding affinity at nanomolar concentration. By ELISA, these binders showed dose-dependent binding to FGFR4 protein but not to other FGFR family members. We then made m410 and m412 into CAR and BiTE format, respectively. T cells transduced with m410 CAR construct were found highly potent in inducing gamma interferon, TNF alpha, and cytotoxicity when the FGFR4-CART are cocultured with RMS cells. Our in vivo testing found them to be effective in eliminating RMS cells in murine xenograft models. When T cells were cocultured with RMS cells in the presence of m412 BiTE in vitro, potent selective antitumor effect was observed, suggesting this can be another promising strategy for RMS immunotherapy. Conclusions: Here our data demonstrated that we had successfully generated binders specific to human FGFR4. The CAR and BiTE developed from these binders were able to kill FGFR4-positive target cells. Our data suggest that these FGFR4 CARs and FGFR4 BiTEs could provide effective immune therapies for rhabdomyosarcoma and other FGFR4-expressing cancers. Citation Format: Adam Cheuk, Nityashree Shivaprasad, Dina Schneider, Marielle Yohe, Meijie Tan, Peter Azorsa, Ronald Sams, Silvia Pomella, Berkley Gryder, Rossella Rota, Ben Stanton, Jun Wei, Young Song, Xinyu Wen, Sivasish Sindiri, Jeetendra Kumar, Robert Hawley, Joon-Yong Chung, Doncho Zhelev, Zhongyu Zhu, Dimiter Dimitrov, Stephen Hewitt, Boro Dropulic, Rimas Orentas, Javed Khan. Development of FGFR4-specific chimeric antibody receptor (CAR) T cell and bispecific T cell engager (BiTE) for rhabdomyosarcoma (RMS) immunotherapy [abstract]. In: Proceedings of the AACR Special Conference on the Advances in Pediatric Cancer Research; 2019 Sep 17-20; Montreal, QC, Canada. Philadelphia (PA): AACR; Cancer Res 2020;80(14 Suppl):Abstract nr A08.

Analysis of related patents on coronavirus monoclonal antibody therapy

<p indent="0mm">Coronaviruses, such as SARS-CoV, MERS-CoV, and the recently epidemic coronavirus (SARS-CoV-2), seriously threaten human health and cause significant economic losses. Monoclonal antibodies targeting coronaviruses may inhibit viral infection by blocking viral binding to the receptor on cells. In this study, the patent applications and clinical trial results for monoclonal antibody drugs against coronavirus were analyzed primarily to provide ideas and references for the application of monoclonal antibodies in the fight against COVID-19, and to accelerate the drug development process. There are currently no established patents related to the monoclonal antibodies against SARS-CoV-2, both due to COVID-19’s relatively recent emergence, and due to the lengthy patent application and publication process. A thorough global analysis of monoclonal antibody patent applications indicates a total of 194 diagnostic monoclonal antibody techniques, accounting for 63%, and 113 therapeutic monoclonal antibody techniques, accounting for 37%. Among those applications related to therapeutic monoclonal antibody technology, product patent applications dominate, with a total number of 96, accounting for 85%. According to the geographical analysis of applicants for coronavirus therapeutic monoclonal antibodies, there are 35 patent applications from, accounting for 31%, and 78 foreign patent applications, accounting for 69%. The analysis of the target of the coronavirus therapeutic monoclonal antibody-related patent applications indicates that there are 61 patent applications for monoclonal antibodies specific to the virus’s own antigen, accounting for 54%. This type of monoclonal antibody specifically targets viral antigens to suppress viruses, whereas the other 52 patent applications in this category target viral non-specific monoclonal antibodies, accounting for 46%. Amongst current studies on the monoclonal antibodies against SARS-CoV and MERS-CoV, relevant patent applications for therapeutic monoclonal antibodies are mainly based on products, and the target is the S protein or its receptor binding region (RBD), which prevents virus infection of cells. Currently, there are no available therapeutic monoclonal antibody drugs against coronavirus. The monoclonal antibody drug specifically targeting the S protein of MERS-CoV is still in clinical trials. Three monoclonal antibody drugs treating for COVID-19 are also in clinical trials, all of which involve second use of the known monoclonal antibodies in their mechanisms. The RBD of SARS-CoV-2 has common-epitope peptide with SARS-CoV. By verifying the effect of the monoclonal antibody against SARS-CoV in the treatment of COVID-19, patent applications for the use of the monoclonal antibody against COVID-19 can be designed. Methods of blocking monoclonal antibodies against the RBD of SARS-CoV-2 can be used as potential drug mechanisms for the treatment of COVID-19. Related patent applications and drug development will also be hot spots in the future.

Uncertainty in an era of transformative therapy for haemophilia: Addressing the unknowns.

Haemophilia is at the dawn of a new era in therapeutic management, one that can generate greater protection from bleeding and a functional cure in some individuals. Prior advances in protein engineering and monoclonal antibody technology have facilitated therapeutic options to maintain decreased risk of bleeding and less burdensome treatment. The use of gene transfer, first proposed in 1971 for monogenic diseases, is emerging as an effective long-term treatment for a variety of diseases. Transfer of functional factor VIII (FVIII) and factor IX (FIX) genes has witnessed a series of advances and setbacks since the first non-clinical experiments in animals were initiated nearly 30years ago. More recently, multiyear therapeutic levels of FVIII and FIX activity have been achieved in human clinical trials, translated into meaningful clinical benefit and a functional cure. While clinical progress has been definitive, many questions remain unanswered as prelicensure phase 3 clinical trials are underway. These unanswered questions translate into a state of uncertainty about the known unknowns and unknown unknowns intrinsic to any new therapeutic platform. Accepting this modality as a means to functionally cure haemophilia also means accepting the uncertainty regarding the biology of viral vector-mediated gene transfer, which remains inadequately understood. Gene therapy is a far more complex biological 'drug' than small molecule and protein drugs, where manufacturing processes and the drugs themselves are now well characterized. Extent of community acceptance of uncertainty and acknowledgement of the need for an uncompromising drive for answers to the unknowns will characterize the introduction of this first generation of gene therapy for haemophilia to the wider patient population in both resource-rich and resource-poor countries.

Modified bacterial outer membrane vesicles induce autoantibodies for tumor therapy

Using monoclonal antibodies to block tumor angiogenesis has yielded effective antitumor effects. However, this treatment method has long cycles and is very expensive; therefore, its long-term and extensive application is limited. In this study, we developed a nanovaccine using bacterial biomembranes as carriers for antitumor therapy. The whole basic fibroblast growth factor (BFGF) molecule (154 amino acids (aa)) was loaded onto bacterial outer membrane vesicles (OMVs) using gene recombination technology. The strong adjuvant effect of OMVs was used to induce the host to produce anti-BFGF autoantibodies. We proved that persistent anti-BFGF autoantibodies can be induced in mice after only 3 immunizations to antagonize BFGF functions. The effects included multiple tumor suppression functions, including inhibition of tumor angiogenesis, induction of tumor cell apoptosis, reversal of tumor immune barriers, and promotion of tumor-specific cytotoxic T lymphocytes (CTLs), eventually causing tumor regression. We confirmed that bacterial biomembranes can be used as a vaccine delivery system to induce the production of antibodies against autoantigens, which may be used for tumor therapy. This study expands the application fields of bacterial biomembrane systems and provides insight for tumor immunotherapy other than monoclonal antibody technology. Statement of SignificanceIn this study, we proved that bacteria-released outer membrane vesicles (OMVs) modified via genetic engineering can be used as a vaccine carrier to break autoimmune tolerance and induce the body to produce autoantibodies to antagonize pathological molecules and block pathological signaling pathways for tumor therapy. OMVs naturally released by bacteria were used to successfully load the full-length BFGF protein (154 aa). We proved that persistent anti-BFGF autoantibodies can be induced in tumor-bearing mice after only 3 immunizations to effectively inhibit tumors. Furthermore, the production of these antibodies successfully inhibited tumor angiogenesis, promoted tumor cell apoptosis, reversed the tumor immunosuppressive microenvironment, increased the cytotoxic T lymphocyte (CTL) reaction, and eventually inhibited tumor growth.