Ana Maria Moro

Instituto Butantan | butanan · Laboratory of Biopharmaceuticals

Tetanus disease, caused by C. tetani, starts with wounds or mucous layer contact. Prevented by vaccination, the lack of booster shots throughout life requires prophylactic treatment in case of accidents. The incidence of tetanus is high in underdeveloped countries, requiring the administration of antitetanus antibodies, usually derived from immuniz...

Tetanus disease is caused by the spore-forming bacteria Clostridium tetani. Its development starts with wounds or mucous layers of spore contact in water, soil, human, and animal feces. It is prevented by vaccination, but the lack of booster shots throughout life and age-decreased immune surveillance by immunosenescence enforces prophylactic treatm...

Introduction Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces rapid production of IgM, IgA, and IgG antibodies directed to multiple viral antigens that may have impact diverse clinical outcomes. Methods We evaluated IgM, IgA, and IgG antibodies directed to the nucleocapsid (NP), IgA and IgG to the Spike protein and...

SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-R...

Disadvantages of using murine monoclonal antibodies (mAb) in human therapy, such as immunogenicity response, led to the development of technologies to transform murine antibodies into human antibodies. The murine anti-FGF2 3F12E7 mAb was proposed as a promising agent to treat metastatic melanoma tumors; once it blocks the FGF2, responsible for play...

Removal of CD4 T cell epitopes from therapeutic antibody sequences is expected to mitigate their potential immunogenicity, but its application is complicated by the location of their T cell epitopes, which mainly overlap with complementarity-determining regions. We therefore evaluated the flexibility of antibody sequences to reduce the predicted af...

Immune responses after COVID-19 vaccination should be evaluated in different populations around the world. This study compared antibody responses induced by ChAdOx1 nCoV-19, CoronaVac, and BNT162b2 vaccines. Blood samples from vaccinees were collected pre- and post-vaccinations with the second and third doses. The study enrolled 78 vaccinees, of wh...

The viruses that cause seasonal Influenza infect between 5-15% of the world's population annually, causing about 650,000 deaths yearly from Severe Acute Respiratory Syndrome (SARS). Every year there is a recurrence of seasonal epidemics caused by the Influenza virus. Vaccination needs annual updating to keep up with viral evolution. Patients at ris...

Tetanus toxin (TeNT) is produced by C. tetani, a spore-forming bacillus broadly spread in the environment. Although an inexpensive and safe vaccine is available, tetanus persists because of a lack of booster shots and variable responses to vaccines due to immunocompromised status or age-decreased immune surveillance. Tetanus is most prevalent in lo...

The COVID-19 pandemic caused by the severe acute syndrome virus 2 (SARS-CoV-2) has been around since November 2019. As of early June 2022, more than 527 million cases were diagnosed, with more than 6.0 million deaths due to this disease. Coronaviruses accumulate mutations and generate greater diversity through recombination when variants with diffe...

Oral tolerance is defined as a specific suppression of cellular and humoral immune responses to a particular antigen through prior oral administration of an antigen. It has unique immunological importance since it is a natural and continuous event driven by external antigens. It is characterized by low levels of IgG in the serum of animals after im...

Hemolytic Uremic Syndrome (HUS) associated with Shiga-toxigenic Escherichia coli (STEC) infections is the principal cause of acute renal injury in pediatric age groups. Shiga toxin type 2 (Stx2) has in vitro cytotoxic effects on kidney cells, including human glomerular endothelial (HGEC) and Vero cells. Neither a licensed vaccine nor effective ther...

Objective To investigate the efficacy and safety of hu3S193, a humanized anti-Lewis-Y monoclonal antibody, as a consolidation strategy in patients with platinum-sensitive recurrent epithelial ovarian cancer who achieved a second complete response after salvage platinum-doublet chemotherapy. Methods This single-arm phase II study accrued patients w...

In the past few years, improvement in computational approaches provided faster and less expensive outcomes on the identification, development, and optimization of monoclonal antibodies (mAbs). In silico methods, such as homology modeling, to predict antibody structures, identification of epitope-paratope interactions, and molecular docking are usef...

Potency testing of tetanus antitoxin must be performed in vivo, in a very painful, stressful and prone to high variability assay. It is, therefore, mandatory to find alternatives to this kind of potency assessment. Immunochemical tests as ELISA or ToBI test are already available but usually results in a poor correlation to the in vivo protection. C...

The use of antibody-based therapy to treat a variety of diseases and conditions is well documented. The use of antibodies as an antidote to treat tetanus infections was one of the first examples of immunotherapy and remains the standard of care for cases involving potential infections. Plasma-derived immunoglobulins obtained from human or horse pos...

Shiga toxin (Stx) producing Escherichia coli (STEC) is responsible for causing hemolytic uremic syndrome (HUS), a life-threatening thrombotic microangiopathy characterized by thrombocytopenia, hemolytic anemia, and acute renal failure after bacterially induced hemorrhagic diarrhea. Until now, there has been neither an effective treatment nor method...

The use of serum containing polyclonal antibodies from animals immunized with toxins marked the beginning of the application of antibody-based therapy in late nineteenth century. Advances in basic research led to the development of the hybridoma technology in 1975. Eleven years later, the first therapeutic monoclonal antibody (mAb) was approved, an...

Early and accurate detection of primary or metastatic tumors is of great value in staging, treatment management, and prognosis. Tumor angiogenesis plays an essential role in the growth, invasion, and metastatic spread of solid cancers, and so, is a promising approach for tumor imaging. The GX1 (CGNSNPKSC) peptide was identified by phage display lib...

Tumor necrosis factor alpha (TNFα) is a pro-inflammatory cytokine that mediates the homeostasis of immune responses; its exacerbated production is associated with the pathogenesis of autoimmune and chronic inflammatory diseases. Anti-TNFα drugs have revolutionized the treatment of inflammatory conditions such as rheumatoid arthritis and Crohn’s dis...

The development and improvement of strategies related to discovery technologies of monoclonal antibodies (mAbs) (phage display, yeast display, ribosome display, bacterial display, mammalian cell surface display, mRNA display, DNA display, transgenic animal, and human B-cell derived) opened perspectives for the screening and the selection of therape...

Stx1 toxin is one of the AB5 toxins of Shiga toxin-producing Escherichia coli (STEC) responsible for foodborne intoxication during outbreaks. The single-chain variable fragment (scFv) is the most common recombinant antibody format; it consists of both variable chains connected by a peptide linker with conserved specificity and affinity for antigen....

e17039 Background: Lewis-Y (LeY) antigen is a blood group related antigen expressed in 75% of OC. hu3S193 is a humanized anti-LeY IgG1 mAb with strong complement and antibody dependent cytotoxicity with clinical benefit shown in a phase II study in pts with platinum-resistant OC and small tumor burden. Improving progression free survival (PFS) in p...

Dependência tecnológica na produção de imunobiológicos no Brasil: transferência de tecnologia versus pesquisa nacional Resumo: O objetivo deste estudo foi analisar as estratégias adotadas para a produção de imunobiológicos no Brasil no ano de 2014/2015 nos principais centros biotecnológicos do país. Foi realizada avaliação de portfólios, artigos ci...

Background and objective: Radiotracer diagnosis of insulinoma, can be done using somatostatin or glucagon-like peptide 1 (GLP-1). Performance of GLP-1 antagonists tends to be better than of agonists. Methods: We investigated the uptake of the antagonist exendin (9-39), radiolabeled with technetium- 99m. Two different sites of the biomolecule wer...

A full copy is available: http://rdcu.be/o3Gf. Genetic characterization of protein-producing clones represents additional value to cell line development. In the present study, ten Per.C6 clones producing a Rebmab100 monoclonal antibody were selected using two cloning methods: six clones originated from limiting dilution cloning and four by the aut...

Gliomas are the most common type among all central nervous system tumors. The aggressiveness of gliomas is correlated with the level of angiogenesis and is often associated with prognosis. The aim of this study is to evaluate the novel GX1 peptide and the heterodimer RGD-GX1 radiolabeled with technetium-99m, for angiogenesis detection in glioma mod...

Compelling evidence suggests that fibroblast growth factor 2 (FGF2), overexpressed in melanomas, plays an important role in tumor growth, angiogenesis and metastasis. In this study, we evaluated the therapeutic use of a new anti-FGF2 monoclonal antibody (mAb), 3F12E7, using for that the B16-F10 melanoma model. The FGF2 neutralizing effect of this a...

The aim of this preclinical study was to evaluate the characteristics of the monoclonal antibody Rebmab200, which is a humanized version of the ovarian-specific murine antibody MX35. This investigation contributes to the foundation for future clinical α-radioimmunotherapy of minimal residual ovarian cancer with 211At-Rebmab200. Here, the biodistrib...

Many patents for the first biologicals derived from recombinant technology and, more recently, monoclonal antibodies (mAbs) are expiring. Naturally, biosimilars are becoming an increasingly important area of interest for the pharmaceutical industry worldwide, not only for emergent countries that need to import biologic products. This review shows t...

Upon infection, pathogenic Leptospira bind several complement regulators in order to overcome host innate immunity. We have previously characterized a 20 kDa leptospiral surface protein which interacts with C4b-binding protein (C4BP), named Leptospiral complement regulator-acquiring protein A (LcpA). Here we show that LcpA also interacts with human...

POSTER PRESENTATION Open Access Strategies for clone detection, selection and isolation in Per.C6 cells - case for Rebmab100 Fernanda P Yeda1,2, Mariana L dos Santos1,2, Lilian R Tsuruta1,2, Bruno B Horta1,2, André L Inocencio1, Oswaldo K Okamoto2,3, Maria C Tuma2, Ana M Moro1* From 23rd European Society for Animal Cell Technology (ESACT) Meeting:...

Victims of massive bee attacks become extremely ill, presenting symptoms ranging from dizziness and headache to acute renal failure and multiple organ failure that can lead to death. Previous attempts to develop specific antivenom to treat these victims have been unsuccessful. We herein report a F(ab)´2-based antivenom raised in horse as a potentia...

NaPi2b, a sodium-dependent phosphate transporter, is highly expressed in ovarian carcinomas and is recognized by the murine monoclonal antibody MX35. The antibody had shown excellent targeting to ovarian cancer in several early phase clinical trials but being murine the antibody's full therapeutic potential could not be explored. To overcome this i...

Digoxin is a pharmaceutical used in the control of cardiac dysfunction. Its therapeutic window is narrow, with effect dosage very close to the toxic dosage. To counteract the toxic effect, polyclonal Fab fragments are commercially available. Our study is based on a monoclonal anti-digoxin antibody, which would provide a product with a specific pote...

Aiming to sort B cells expressing anti-tetanus toxoid IgG from human blood that could be used for heavy and light chain amplification and cloning, this work started by the preliminary detection of such cells in 10 mL of human peripheral blood. After informed consent, blood was drawn from six volunteers and the mononuclear cells (PBMC) were stained...

Impaired activity of the lysosomal enzyme glucocerebrosidase (GCR) results in the inherited metabolic disorder known as Gaucher disease. Current treatment consists of enzyme replacement therapy by administration of exogenous GCR. Although effective, it is exceptionally expensive, and patients worldwide have a limited access to this medicine. In Bra...

Scintigraphy is generally not the first choice treatment for prostate cancer, although successful studies using bombesin analog radiopeptides have been performed. Recently, a novel peptide obtained using a phage display library demonstrated an affinity for prostate tumor cells. The aim of this study was to compare the use of a bombesin analog to th...

Early and reliable diagnosis of melanoma, a skin tumor with a poor prognosis, is extremely important. Phage display peptide libraries are a convenient screening resource for identifying bioactive peptides that interact with cancer targets. The aim of this study was to evaluate two technetium-99m tracers for angiogenesis detection in a melanoma mode...

Cancer has been investigated using various pre-targeting techniques or models focusing on radiobombesin analogues; however, both are not offered together. In this study, nano-bombesin labeling by a pre-targeting system was undertaken to develop an alternative approach for prostate tumor treatment. A two-step pre-targeting system utilizing a combina...

Two humanized monoclonal antibody constructs bearing the same variable regions of an anti-CD3 monoclonal antibody, whole IgG and FvFc, were expressed in CHO cells. Random and site-specific integration were used resulting in similar expression levels. The transfectants were selected with appropriate selection agent, and the surviving cells were plat...

Strategies to minimize the immunogenicity and toxicity of murine anti-CD3 antibodies (e.g. OKT3) are of special interest for organ transplantation and for the treatment of autoimmune diseases. In the present work, we have developed two humanized anti-CD3 antibodies. These molecules were shown to bind to human CD3, though less efficiently, and displ...

Chitosan, alpha-(1-4)-amino-2-deoxy-beta-D-glucan, is a deacetylated form of chitin, an abundant natural polysaccharide present in crustacean shells. Its unique characteristics such as positive charge, biodegradability, biocompatibility, nontoxicity, and rigid linear molecular structure make this macromolecule ideal as drug carrier. The association...

In this work the closest human germline sequence was used as the framework on to which to graft murine CDRs. The proposed fully humanized version displays the same staining pattern to different cell subpopulations like CD4, CD8 and memory CD45RO lymphocytes, when compared with the original anti-CD18 MAb, reinforcing the germline approach as a succe...

In this work the closest human germline sequence was used as the framework on to which to graft murine CDRs. The proposed fully humanized version displays the same staining pattern to different cell subpopulations like CD4, CD8 and memory CD45RO lymphocytes, when compared with the original anti-CD18 MAb, reinforcing the germline approach as a succe...

Objetivos: O anticorpo monoclonal anti-CD3 tem sido empregado na prevenção e tratamento de episódios de rejeição aguda em transplante de órgãos. Neste estudo retrospectivo, relatamos a experiência de três instituições brasileiras que utilizaram o anticorpo anti-CD3 produzido pelo Instituto Butantan (São Paulo, Brasil) em pacientes transplantados re...

Purposes: The monoclonal anti-CD3 antibody has been used as induction and to treat acute rejection episodes in organ transplantation. This retrospective study reports a trial in three Brazilian transplantation centers, which have used a national anti-CD3 preparation developed by the Butantan Institute (São Paulo, Brazil), in renal transplant patien...

Microspheres of polymers like poly(lactic-co-glycolic acid) (PLGA) have been studied as a vehicle for controlled release vaccines. They require materials and processes that might change the protein antigenicity. Lactic acid is produced during microsphere degradation that occurs in tandem with protein liberation. In addition, most of the proteins th...

Humanization of monoclonal antibodies by complementary determinant region (CDR)-grafting has become a standard procedure to improve the clinical usage of animal antibodies. However, antibody humanization may result in loss of activity that has been attributed to structural constraints in the framework structure. In this paper, we report the complet...

The traditional way for production of recombinant human erythropoietin (rhEPO) is through cultivation of the clone in roller bottles when the cells are adherence dependent. This system was used in this study, aiming to compare different categories of fetal bovine sera for media supplementation as well as two serum free media for the production or r...

Erythropoietin (EPO) is a glycoprotein hormone responsible for the maintenance of red blood cells number during steady-state condidons and for accelerating recovery of red blood cell mass following hemorrhage. The primary stimulus for increased EPO synthesis is tissue hypoxia resulting from decreased blood O2 availability. The principal function of...

The 6.7 murine monoclonal antibody (mAb) recognizes the human CD18 antigen and is therefore of interest as an anti-inflammatory agent. The 6.7 heavy variable chain (VH) was humanized using the closest human germline sequence as the template on to which to graft the murine complementary determining regions (CDRs). Two versions were proposed, one in...

Hybridoma cells can be cultivated at high densities for long periods through the combination of immobilization devices and continuous perfusion of medium, providing an efficient oxygen and nutrients feeding without cell loss. The disadvantage of these systems is that the cell density cannot be directly monitored and the cellular health is followed...

The immobilisation of cells in a perfusion culture allows to obtain a high cell concentration and an efficient removal of the catabolites without cell loss. A disadvantage of this system is that the cell density cannot be directly monitored. The cellular metabolism is just followed by online measurements of pH and dissolved oxygen (DO) and off-line...

One of the main concerns about therapeutic products derived from mammalian cell cultures is the residual DNA content. The maximum DNA content per dose is established as 100 pg for European and American authorities. For monoclonal antibody production we have been cultivating hybridomas in a packed-bed bioreactor with internal aeration with continuou...

Besides surgery, the therapeutic possibilities for the treatment of human gliomas include adoptive cellular immunotherapy, radioimmunotherapy, immunotherapy mediated by chemoimmunoconjugates and, more recently, bispecific monoclonal antibodies (biMAbs). Anti-CD3 x anti-tenascin (TN) is the first reagent of a number of biMAbs under investigation for...

Monoclonal antibodies are one of the most important products of biotechnology and laboratories and companies all over the world are pursuing their large-scale production. Herein we report a protocol for hybridoma cell cultivation over small glass cylinders inside a 3 liter bioreactor vessel which leads to the production and purification--in order o...

A highly significant familial aggregation of eosinophil levels ( X 2/3 = 38.00) was detected in a sample from three Brazilian populations with a high incidence of helminthic parasitism. The data were unable to resolve genetic or common environment causation due to the lack of environmental concomitant variables. Results of path analysis indicate th...

Recently, a significant sex distortion of the esterase D locus segregation, both in Portugal and Germany, was reported. In contrast to these puzzling results, data on around 1,200 informative northeastern Brazilian individuals did not show any signs of a sex-phenotype association (chi 2 = 0.35). The joint analysis of this sample together with that...

1. Phosphoserine phosphatase (PSP), specific for D-and L-phosphoserine, has been identified in all human tissues. 2. PSP shows no activity towards phosphorylated serine residues in phosphoproteins. 3. In most tissues, PSP consists of a single major isozyme, probably determined by a single autosomal locus. 4. Other isozymes, identified quite frequen...

The frequency of cells presenting chromosome abnormalities was determined in patients with schistosomiasis before and after treatment with a single dose of 2.5 mg/kg body weight of hycanthone methanosulfonate. No significant effect of the drug was detected with this therapeutic dose.