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Molecular & Cellular Proteomics : MCP 2021Antibodies play essential roles in both diagnostics and therapeutics. Epitope mapping is essential to understand how an antibody works and to protect intellectual...
Antibodies play essential roles in both diagnostics and therapeutics. Epitope mapping is essential to understand how an antibody works and to protect intellectual property. Given the millions of antibodies for which epitope information is lacking, there is a need for high-throughput epitope mapping. To address this, we developed a strategy, Antibody binding epitope Mapping (AbMap), by combining a phage displayed peptide library with next-generation sequencing. Using AbMap, profiles of the peptides bound by 202 antibodies were determined in a single test, and linear epitopes were identified for >50% of the antibodies. Using spike protein (S1 and S2)-enriched antibodies from the convalescent serum of one COVID-19 patient as the input, both linear and potentially conformational epitopes of spike protein specific antibodies were identified. We defined peptide-binding profile of an antibody as the binding capacity (BiC). Conceptually, the BiC could serve as a systematic and functional descriptor of any antibody. Requiring at least one order of magnitude less time and money to map linear epitopes than traditional technologies, AbMap allows for high-throughput epitope mapping and creates many possibilities.
Topics: Antibodies, Monoclonal; Antibodies, Viral; COVID-19; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Epitopes; Escherichia coli Proteins; High-Throughput Nucleotide Sequencing; Humans; Immune Sera; Peptide Library; Spike Glycoprotein, Coronavirus
PubMed: 33109704
DOI: 10.1074/mcp.RA120.002314 -
Transfusion and Apheresis Science :... Aug 2020Passive immunotherapy with plasma derived from patients convalescent from SARS-CoV-2 infection can be a promising approach in the treatment of COVID-19 patients. It is... (Review)
Review
Passive immunotherapy with plasma derived from patients convalescent from SARS-CoV-2 infection can be a promising approach in the treatment of COVID-19 patients. It is important that Blood Establishments are prepared to satisfy requests for immune plasma by defining the requirements applicable to plasma donors and the standards for preparation, qualification, storage, distribution and control of use of the product. This position paper is aimed to give recommendations on biological characteristics of a plasma preparation from convalescent donors and to support the evaluation of this therapeutic approach in more rigorous investigations.
Topics: Antibodies, Viral; Betacoronavirus; Blood Component Removal; Blood Donors; COVID-19; Coronavirus Infections; Donor Selection; Humans; Immune Sera; Immunization, Passive; Pandemics; Pneumonia, Viral; Product Labeling; Risk; SARS-CoV-2; COVID-19 Serotherapy
PubMed: 32532691
DOI: 10.1016/j.transci.2020.102817 -
The Journal of Urology Feb 2017Rabbit antiserum raised against the crude extract of normal human prostatic tissue contained antibodies to a prostatic tissue-specific antigen as shown by...
Rabbit antiserum raised against the crude extract of normal human prostatic tissue contained antibodies to a prostatic tissue-specific antigen as shown by immunoprecipitation techniques. Using this antiserum a prostate antigen was detected in normal, benign hypertrophic, and malignant prostatic tissues, but not in other human tissues. The prostate antigen was purified to homogeneity from prostatic tissues and showed a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing. This report thus presents the first demonstration of the purification of a prostate-specific antigen that does not represent prostatic acid phosphatase.
Topics: Animals; Electrophoresis, Polyacrylamide Gel; Female; Humans; Immune Sera; Immunoprecipitation; Isoelectric Focusing; Kallikreins; Male; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Rabbits
PubMed: 28012750
DOI: 10.1016/j.juro.2016.10.100 -
JAMA May 2020
Topics: History, 20th Century; Humans; Immune Sera; Influenza Pandemic, 1918-1919; Influenza Vaccines; Influenza, Human
PubMed: 32396175
DOI: 10.1001/jama.2020.6524 -
Glycoconjugate Journal Oct 2019Even though a vaccine that targets tumor-associated carbohydrate antigens on epithelial carcinoma cells presents an attractive therapeutic approach, relatively poor...
Even though a vaccine that targets tumor-associated carbohydrate antigens on epithelial carcinoma cells presents an attractive therapeutic approach, relatively poor immunogenicity limits its development. In this study, we investigated the immunological activity of a fluoro-substituted Sialyl-Tn (F-STn) analogue coupled to the non-toxic cross-reactive material of diphtheria toxin197 (CRM197). Our results indicate that F-STn-CRM197 promotes a greater immunogenicity than non-fluorinated STn-CRM197. In the presence or absence of adjuvant, F-STn-CRM197 remarkably enhances both cellular and humoral immunity against STn by increasing antigen-specific lymphocyte proliferation and inducing a mixed Th1/Th2 response leading to production of IFN-γ and IL-4 cytokines, as well as STn-specific antibodies. Furthermore, antisera produced from F-STn-CRM197 immunization significantly recognizes STn-positive tumor cells and increases cancer cell lysis induced by antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) pathways. Our data suggest that this F-STn vaccine may be useful for cancer immunotherapy and possibly for prophylactic prevention of cancer.
Topics: Adjuvants, Immunologic; Animals; Antibodies, Neoplasm; Antibody-Dependent Cell Cytotoxicity; Antigens, Tumor-Associated, Carbohydrate; Bacterial Proteins; Cancer Vaccines; Cell Line, Tumor; Colonic Neoplasms; Female; Gene Expression; Glycoconjugates; Halogenation; Humans; Immune Sera; Immunity, Cellular; Immunity, Humoral; Immunization; Immunogenicity, Vaccine; Interferon-gamma; Interleukin-4; Lymphocytes; Mice; Mice, Inbred BALB C; Spleen; Th1-Th2 Balance
PubMed: 31267246
DOI: 10.1007/s10719-019-09884-0 -
PloS One 2023Influenza B viruses (IBV) are responsible for a considerable part of the burden caused by influenza virus infections. Since their emergence in the 1980s, the Yamagata...
Influenza B viruses (IBV) are responsible for a considerable part of the burden caused by influenza virus infections. Since their emergence in the 1980s, the Yamagata and Victoria antigenic lineages of influenza B circulate in alternate patterns across the globe. Furthermore, their evolutionary divergence and the appearance of new IBV subclades complicates the prediction of future influenza vaccines compositions. It has been proposed that the addition of the neuraminidase (NA) antigen could potentially induce a broader protection and compensate for hemagglutinin (HA) mismatches in the current vaccines. Here we show that anti-NA and -HA sera against both Victoria and Yamagata lineages have limited inter-lineage cross-reactivity. When transferred to mice prior to infection with a panel of IBVs, anti-NA sera were as potent as anti-HA sera in conferring protection against homologous challenge and, in some cases, conferred superior protection against challenge with heterologous IBV strains.
Topics: Animals; Humans; Mice; Antibodies, Viral; Cross Protection; Hemagglutinin Glycoproteins, Influenza Virus; Hemagglutinins; Immune Sera; Influenza B virus; Influenza, Human; Neuraminidase; Orthomyxoviridae Infections
PubMed: 36689429
DOI: 10.1371/journal.pone.0280825 -
Animal Biotechnology Dec 2023Viperin, also known as radical S-adenosyl methionine domain-containing protein (RSAD2) is a multifunctional interferon-stimulated gene (ISG) that is activated during the...
Viperin, also known as radical S-adenosyl methionine domain-containing protein (RSAD2) is a multifunctional interferon-stimulated gene (ISG) that is activated during the viral infections. Viperin belongs to S-adenosyl methionine (SAM) superfamily of enzymes known to catalyze radical-mediated reactions and viperin inhibits a wide range of DNA and RNA viruses through its broad range of activity. The present study reports cloning and expression of bovine viperin in a bacterial expression system. PCR-based site-directed mutagenesis was carried out for deletion of N-terminal 1-70 amino acid containing amphipathic helix of viperin that interferes in protein expression and purification. The resultant truncated viperin protein was expressed in , BL-21(DE3) competent cells and purified using nickel charged affinity column. The truncated 54 kDa protein was confirmed by western blot using human RSAD2 as a probe. Further, in house, hyperimmune serum was raised against the truncated viperin in the rabbit and the reactivity was confirmed by western blot using mammalian expression vector construct of viperin transfected in Baby Hamster kidney (BHK) cells and in MDBK cells infected with Foot and Mouth disease Asia I virus.
Topics: Animals; Cattle; Humans; Rabbits; Immune Sera; Proteins; Methionine; Mammals
PubMed: 36112063
DOI: 10.1080/10495398.2022.2120890 -
Journal of Infection in Developing... May 2022Diverse serogroups of Escherichia coli cause sporadic cases and outbreaks of diarrhea among children. Our study aimed to evaluate the serogroups of diarrheagenic strains...
INTRODUCTION
Diverse serogroups of Escherichia coli cause sporadic cases and outbreaks of diarrhea among children. Our study aimed to evaluate the serogroups of diarrheagenic strains of E. coli that cause diarrheal disease in children under two years old, and clarify if the cases were sporadic or outbreaks.
METHODOLOGY
The retrospective study included 130 strains of pathogenic E. coli, isolated from children who were less than two years of age, and had diarrheal disease, between May 2016 and July 2019. The study was conducted in the Bacteriology Laboratory (County Clinical Hospital, Mureș, Romania). The 130 strains were sero-grouped using polyvalent and monovalent O antisera. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) was performed to evaluate the similarity between different E. coli strains, and a simplex polymerase chain reaction (PCR) was performed to detect the presence of the hlyA gene that is specific to the enterohemorrhagic strains.
RESULTS
After agglutination with polyvalent O antisera, slightly more than half of the strains (50.77%) were sero-grouped as Shiga toxin-producing E. coli (STEC), and the rest of the strains belonged to the Enteropathogenic Escherichia coli (EPEC) serogroups. Serogroup O157 was the most frequently identified (16.51% of the total number of typeable strains), and one strain was positive for hlyA. ERIC-PCR revealed a high diversity of strains, with an overall 50% similarity.
CONCLUSIONS
STEC serogroups were the most common strains causing diarrheal disease, and O-157 was the dominant serogroup identified. The strains included in our study presented high genetic diversity, suggesting that most of the cases were sporadic.
Topics: Child; Diarrhea; Enteropathogenic Escherichia coli; Escherichia coli Infections; Genetic Variation; Humans; Immune Sera; Infant; Retrospective Studies; Serogroup
PubMed: 35656954
DOI: 10.3855/jidc.15703 -
Toxicon : Official Journal of the... Apr 2020Accidents by freshwater stingrays are common in northern Brazil, there is no specific therapy for high morbidity and local tissue destruction. The irradiation of venoms...
Accidents by freshwater stingrays are common in northern Brazil, there is no specific therapy for high morbidity and local tissue destruction. The irradiation of venoms and toxins by ionizing radiation has been used to produce appropriate immunogens for the production of antisera. We planned to study the efficacy of stinging mucus irradiation in the production of antisera, with serum neutralization assays of edematogenic activity and quantification of cytokines performed in animal models of immunization with native and irradiated mucus of Paratrygon aiereba, a large freshwater stingray. Antiserum potency and its cross-reactivity with mucus from other freshwater stingrays were detected by ELISA. Immunization models demonstrated the ability to stimulate a strong humoral response with elevated levels of serum IgG detectable by ELISA, and both native and irradiated mucus were immunogenic and capable of recognizing mucus proteins from other freshwater neotropical stingrays. Mucus P. aiereba causes cellular and humoral adaptive immune responses in cells of immunized mice producing antibodies and cytokines such as TNF-α, IL-6 and IL-17. Rabbit antisera immunized with mucus from P. aiereba irradiated at 2 kGy showed a significant reduction of mucus-induced edematogenic activity in mice. Our data suggest that the use of antisera against freshwater stingray mucus show the possibility of specific therapy for these accidents.
Topics: Animals; Bites and Stings; Brazil; Edema; Elasmobranchii; Enzyme-Linked Immunosorbent Assay; Fresh Water; Immune Sera; Mice; Models, Theoretical; Mucus; Pain; Rabbits; Skates, Fish
PubMed: 32067999
DOI: 10.1016/j.toxicon.2020.02.012 -
Molecular and Cellular Endocrinology Jan 2017Irisin, the product of fibronectin type III domain-containing protein 5 (FNDC5) gene, is well-documented to be a regulator of energy metabolism. At present, not much is...
Irisin, the product of fibronectin type III domain-containing protein 5 (FNDC5) gene, is well-documented to be a regulator of energy metabolism. At present, not much is known about its biological function in non-mammalian species. In this study, a full-length tilapia FDNC5 was cloned and its tissue expression pattern has been confirmed. Based on the sequence obtained, we produced and purified recombinant irisin which could induce uncoupling protein 1 (UCP1) gene expression in tilapia hepatocytes. Further, the rabbit polyclonal irisin antiserum was produced and its specificity was confirmed by antiserum preabsorption. In tilapia pituitary cells, irisin inhibited growth hormone (GH) gene expression and secretion and triggered rapid phosphorylation of Akt, Erk1/2, and p38 MAPK. Furthermore, irisin-inhibited GH mRNA expression could be prevented by inhibiting PI3K/Akt, MEK1/2, and p38 MAPK, respectively. Apparently, fish irisin can act directly at the pituitary level to inhibit GH transcript expression via multiple signaling pathways.
Topics: Amino Acid Sequence; Animals; Base Sequence; Biological Assay; Cells, Cultured; Cloning, Molecular; DNA, Complementary; Fibronectins; Gene Expression Profiling; Gene Expression Regulation; Growth Hormone; Humans; Immune Sera; MAP Kinase Signaling System; Pituitary Gland; Recombinant Proteins; Sequence Alignment; Tilapia; Uncoupling Protein 1
PubMed: 27693813
DOI: 10.1016/j.mce.2016.09.030