scholarly journals Specific and Generic Immunorecognition of Glycopeptide Antibiotics Promoted by Unique and Multiple Orientations of Hapten

Biosensors ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 52 ◽  
Author(s):  
Maksim Burkin ◽  
Inna Galvidis ◽  
Sergei Eremin
Keyword(s):  
Immune Responses ◽  
Dynamic Range ◽  
Polyclonal Antibodies ◽  
Recognition System ◽  
Cross Reactivity ◽  
Glycopeptide Antibiotics ◽  
Ic50 Values ◽  
Conjugation Chemistry ◽  
Heterologous Antigens ◽  
Selection Of

Conjugation chemistry does not always provide adequate spatial orientation of hapten in immunogens for the best presentation of generic or individual epitopes. In the present study, the influence of unique and multiple orientations of immunizing hapten on the immune response repertoire was compared to select generic recognition system. The glycopeptides, teicoplanin (TPL) and ristomycin (RSM), were conjugated to BSA to produce immunogens with unique and multiple orientations of haptens. Polyclonal antibodies generated against TPL conjugated through a single site were of uniform specificity and demonstrated selective TPL recognition, regardless of the coating conjugates design. The sensitivity (IC50) of 4 enzyme-linked immunosorbent assays (ELISAs) for TPL varied little within the 3.5–7.4 ng/mL, with a dynamic range of 0.2–100 ng/mL. RSM was coupled to BSA through several glycoside sites that evoked a wider repertoire of response. This first described anti-RSM antibody was selective for RSM in homologous hapten-coated ELISAs with IC50 values in the range 4.2–35 ng/mL. Among the heterologous antigens, periodate-oxidized TPL conjugated to gelatine was selected as the best binder of generic anti-RSM fraction. The developed ELISA showed group recognition of glycopeptides RSM, TPL, eremomycin, and vancomycin with cross-reactivity of 37–100% and a 10–10,000 ng/mL dynamic range. Thus, multiple presentations of immunizing hapten help expand the repertoire of immune responses and opportunities for the selection of the required fine-specificity agent.

scholarly journals Identification of Two Distinct Types of Flagellar Cap Proteins, FliD, in Pseudomonas aeruginosa

2000 ◽  
Vol 68 (3) ◽  
pp. 1474-1479 ◽  
Author(s):  
Shiwani K. Arora ◽  
Nandini Dasgupta ◽  
Stephen Lory ◽  
Reuben Ramphal
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Polyclonal Antibodies ◽  
Immunoblot Analysis ◽  
Protein Product ◽  
Cross Reactivity ◽  
Nucleotide Sequencing ◽  
Host Immune System ◽  
Previous Infection ◽  
Flagellar Proteins ◽  
Selection Of

ABSTRACT Binding of Pseudomonas aeruginosa strain PAK to mucin has been shown to be mediated by the flagellar cap protein, product of the fliD gene. Since the flagellar cap is very likely an exposed structure, the FliD polypeptide should be recognized by the host immune system, analogous to the recognition of dominant epitopes located in the exposed parts of the flagellin polypeptide within the assembled flagellum. In P. aeruginosa, a number of distinct flagellin variants are made, and these variable sequences presumably allow the newly infected P. aeruginosa to escape recognition by the antibody induced during a previous infection. Since similar mechanisms may direct the selection of FliD variants, we examined the extent of sequence heterogeneity among various FliD sequences among a selected group of P. aeruginosa. The results of PCR and nucleotide sequencing of the fliD region of eight different P. aeruginosa strains (laboratory strains PAK, PAO1, and PA103; clinical strains 1244, CS2, and CS32; cystic fibrosis strains CS29 and MDR) suggested that there were two distinct types of FliD in P. aeruginosa, which we named A type and B type. The results of Western blotting using the polyclonal antibodies raised against the purified FliD of A type (PAK) or B type (PAO1) further confirmed the existence of two distinct antigenic types of FliD proteins, with no cross-reactivity between the two serotypes. Further Western immunoblot analysis of the same strains using polyclonal FliC antibody showed that the strains with A-type FliD possessed a-type FliC and those with B-type FliD had b-type FliC. Similar Western blot analyses of 50 more P. aeruginosastrains obtained from varied sources revealed that all strains contained either A-type or B-type FliD, suggesting the existence of only two types of FliD in P. aeruginosa and indicating thatfliC and fliD were coinherited. This limited diversity of FliC and FliD serotypes seems to be a unique feature of flagellar proteins. A chromosomal mutant having an insertion in thefliD gene of P. aeruginosa PAO1 was constructed. The motility defect of this mutant and a previously constructed PAK fliD mutant was better complemented with the fliD gene of the homologous types.

Synthesis and Identification of Norfloxacin Artificial Antigen

2012 ◽  
Vol 433-440 ◽  
pp. 697-702 ◽  
Author(s):  
Jun Wei Liu ◽  
Jin Qing Jiang ◽  
Hai Tang Zhang ◽  
Guo Ying Fan ◽  
Zhi Xing An
Keyword(s):  
Dynamic Range ◽  
Food Samples ◽  
Cross Reactivity ◽  
Assay Performance ◽  
Ic50 Value ◽  
Artificial Antigen ◽  
Sds Page ◽  
Visible Spectra ◽  
Ic50 Values

A multiresidue immunoassay method for determination of Fluoroquinolones (FQs) residues has been developed. For this purpose, NHS ester technology was employed to synthesize the immunogen and coating antigen of Norfloxacin (NFLX). SDS-PAGE, UV-visible spectra and Infrared spectra identification showed that the artificial antigen was conjugated successfully. Based on the square matrix titration, an icELISA method was established. The dynamic range in assay buffer was from 0.038 to 112.8 ng/mL, with LOD and IC50 value of 0.02 ng/mL and 1.2 ng/mL, respectively. This assay showed a high cross-reactivity to Ciprofloxacin (86%), Enrofloxacin (75%), Difloxacin (63%), Sarafloxacin (57%) and Pefloxacin (33.8%). The chemical effects on assay performance showed that the physiological pH (7.4) in assay buffer pursued the maximum absorbance (Amax) and the most sensitive IC50 values. The results suggest the artificial antigen was synthesized successfully, and the established immunoassay could be used for simultaneous detecting of Norfloxacin, Ciprofloxacin, Enrofloxacin, Difloxacin, Sarafloxacin and Pefloxacin residues in animal-original food samples.

scholarly journals From Anti-EBV Immune Responses to the EBV Diseasome via Cross-reactivity

2020 ◽  
Vol 07 (02) ◽  
pp. 051-063
Author(s):  
Darja Kanduc ◽  
Yehuda Shoenfeld
Keyword(s):  
Immune Responses ◽  
Negative Selection ◽  
Epstein Barr Virus ◽  
Cross Reactivity ◽  
Sequence Analyses ◽  
Barr Virus ◽  
Human Proteins ◽  
Epstein Barr ◽  
Reactive Lymphocytes ◽  
Selection Of

AbstractSequence analyses highlight a massive peptide sharing between immunoreactive Epstein-Barr virus (EBV) epitopes and human proteins that—when mutated, deficient or improperly functioning—associate with tumorigenesis, diabetes, lupus, multiple sclerosis, rheumatoid arthritis, and immunodeficiencies, among others. Peptide commonality appears to be the molecular platform capable of linking EBV infection to the vast EBV-associated diseasome via cross-reactivity and questions the hypothesis of the “negative selection” of self-reactive lymphocytes. Of utmost importance, this study warns that using entire antigens in anti-EBV immunotherapies can associate with autoimmune manifestations and further supports the concept of peptide uniqueness for designing safe and effective anti-EBV immunotherapies.

A Study of Different Methodologies Helpful in the Identification of Offline Handwritten Script

2018 ◽  
Vol 6 (6) ◽  
pp. 307
Author(s):  
Manish M. Kayasth ◽  
Bharat C. Patel
Keyword(s):  
Feature Extraction ◽  
Character Recognition ◽  
Recognition Rate ◽  
Recognition System ◽  
Post Processing ◽  
Classification Technique ◽  
Scanned Image ◽  
Gujarati Language ◽  
High Degree ◽  
Selection Of

The entire character recognition system is logically characterized into different sections like Scanning, Pre-processing, Classification, Processing, and Post-processing. In the targeted system, the scanned image is first passed through pre-processing modules then feature extraction, classification in order to achieve a high recognition rate. This paper describes mainly on Feature extraction and Classification technique. These are the methodologies which play an important role to identify offline handwritten characters specifically in Gujarati language. Feature extraction provides methods with the help of which characters can identify uniquely and with high degree of accuracy. Feature extraction helps to find the shape contained in the pattern. Several techniques are available for feature extraction and classification, however the selection of an appropriate technique based on its input decides the degree of accuracy of recognition. 

Elaboration of Novel TTK1 Inhibitory Leads via QSAR-Guided Selection of Crystallographic Pharmacophores Followed By In vitro Assay

2020 ◽  
Vol 16 ◽  
Author(s):  
Mahmoud A. Al-Sha'er ◽  
Mutasem O. Taha
Keyword(s):  
Standard Error ◽  
Qsar Model ◽  
Qsar Modeling ◽  
Vitro Assay ◽  
Physicochemical Descriptors ◽  
Qsar Models ◽  
Ic50 Values ◽  
Pharmacophore Hypotheses ◽  
Selection Of

Introduction: Tyrosine threonine kinase (TTK1) is a key regulator of chromosome segregation. TTK targeting received recent concern for the enhancement of possible anticancer therapies. Objective: In this regard we employed our well-known method of QSAR-guided selection of best crystallographic pharmacophore(s) to discover considerable binding interactions that anchore inhibitors into TTK1 binding site. Method:Sixtyone TTK1 crystallographic complexes were used to extract 315 pharmacophore hypotheses. QSAR modeling was subsequently used to choose a single crystallographic pharmacophore that when combined with other physicochemical descriptors elucidates bioactivity discrepancy within a list of 55 miscellaneous inhibitors. Results: The best QSAR model was robust and predictive (r2(55) = 0.75, r2LOO = 0.72 , r2press against external testing list of 12 compounds = 0.67), Standard error of estimate (training set) (S)= 0.63 , Standard error of estimate (testing set)(Stest) = 0.62. The resulting pharmacophore and QSAR models were used to scan the National Cancer Institute (NCI) database for new TTK1 inhibitors. Conclusion: Five hits confirmed significant TTK1 inhibitory profiles with IC50 values ranging between 11.7 and 76.6 micM.

scholarly journals Perspectives for improvement of Mycoplasma hyopneumoniae vaccines in pigs

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Dominiek Maes ◽  
Filip Boyen ◽  
Bert Devriendt ◽  
Peter Kuhnert ◽  
Artur Summerfield ◽  
...  
Keyword(s):  
Immune Responses ◽  
Respiratory Infections ◽  
Mycoplasma Hyopneumoniae ◽  
Administration Route ◽  
Limited Effect ◽  
Partial Protection ◽  
Carrier Molecule ◽  
Humoral Responses ◽  
Porcine Respiratory ◽  
Selection Of

AbstractMycoplasma hyopneumoniae (M. hyopneumoniae) is one of the primary agents involved in the porcine respiratory disease complex, economically one of the most important diseases in pigs worldwide. The pathogen adheres to the ciliated epithelium of the trachea, bronchi, and bronchioles, causes damage to the mucosal clearance system, modulates the immune system and renders the animal more susceptible to other respiratory infections. The pathogenesis is very complex and not yet fully understood. Cell-mediated and likely also mucosal humoral responses are considered important for protection, although infected animals are not able to rapidly clear the pathogen from the respiratory tract. Vaccination is frequently practiced worldwide to control M. hyopneumoniae infections and the associated performance losses, animal welfare issues, and treatment costs. Commercial vaccines are mostly bacterins that are administered intramuscularly. However, the commercial vaccines provide only partial protection, they do not prevent infection and have a limited effect on transmission. Therefore, there is a need for novel vaccines that confer a better protection. The present paper gives a short overview of the pathogenesis and immune responses following M. hyopneumoniae infection, outlines the major limitations of the commercial vaccines and reviews the different experimental M. hyopneumoniae vaccines that have been developed and tested in mice and pigs. Most experimental subunit, DNA and vector vaccines are based on the P97 adhesin or other factors that are important for pathogen survival and pathogenesis. Other studies focused on bacterins combined with novel adjuvants. Very few efforts have been directed towards the development of attenuated vaccines, although such vaccines may have great potential. As cell-mediated and likely also humoral mucosal responses are important for protection, new vaccines should aim to target these arms of the immune response. The selection of proper antigens, administration route and type of adjuvant and carrier molecule is essential for success. Also practical aspects, such as cost of the vaccine, ease of production, transport and administration, and possible combination with vaccines against other porcine pathogens, are important. Possible avenues for further research to develop better vaccines and to achieve a more sustainable control of M. hyopneumoniae infections are discussed.

scholarly journals Examination of the Efficacy and Cross-Reactivity of a Novel Polyclonal Antibody Targeting the Disintegrin Domain in SVMPs to Neutralize Snake Venom

Toxins ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 254
Author(s):  
Shelby S. Szteiter ◽  
Ilse N. Diego ◽  
Jonathan Ortegon ◽  
Eliana Salinas ◽  
Abcde Cirilo ◽  
...  
Keyword(s):  
Snake Venom ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Cross Reactivity ◽  
Snake Envenomation ◽  
Disintegrin Domain ◽  
The Common ◽  
Neutralization Ability ◽  
New Strategies

Snake envenomation can result in hemorrhage, local necrosis, swelling, and if not treated properly can lead to adverse systemic effects such as coagulopathy, nephrotoxicity, neurotoxicity, and cardiotoxicity, which can result in death. As such, snake venom metalloproteinases (SVMPs) and disintegrins are two toxic components that contribute to hemorrhage and interfere with the hemostatic system. Administration of a commercial antivenom is the common antidote to treat snake envenomation, but the high-cost, lack of efficacy, side effects, and limited availability, necessitates the development of new strategies and approaches for therapeutic treatments. Herein, we describe the neutralization ability of anti-disintegrin polyclonal antibody on the activities of isolated disintegrins, P-II/P-III SVMPs, and crude venoms. Our results show disintegrin activity on platelet aggregation in whole blood and the migration of the SK-Mel-28 cells that can be neutralized with anti-disintegrin polyclonal antibody. We characterized a SVMP and found that anti-disintegrin was also able to inhibit its activity in an in vitro proteolytic assay. Moreover, we found that anti-disintegrin could neutralize the proteolytic and hemorrhagic activities from crude Crotalus atrox venom. Our results suggest that anti-disintegrin polyclonal antibodies have the potential for a targeted approach to neutralize SVMPs in the treatment of snakebite envenomations.

scholarly journals Development of Neutralizing Monoclonal Antibodies for Oncogenic Human Papillomavirus Types 31, 33, 45, 52, and 58

2014 ◽  
Vol 21 (4) ◽  
pp. 587-593 ◽  
Author(s):  
Martha J. Brown ◽  
Hanna Seitz ◽  
Victoria Towne ◽  
Martin Müller ◽  
Adam C. Finnefrock
Keyword(s):  
Monoclonal Antibodies ◽  
Human Papillomavirus ◽  
Immune Responses ◽  
Neutralizing Antibodies ◽  
Cervical Cancers ◽  
Hpv Types ◽  
Oncogenic Hpv ◽  
Neutralizing Epitopes ◽  
Selection Of

ABSTRACTHuman papillomavirus (HPV) is the etiological agent for all cervical cancers, a significant number of other anogenital cancers, and a growing number of head and neck cancers. Two licensed vaccines offer protection against the most prevalent oncogenic types, 16 and 18, responsible for approximately 70% of cervical cancer cases worldwide and one of these also offers protection against types 6 and 11, responsible for 90% of genital warts. The vaccines are comprised of recombinantly expressed major capsid proteins that self-assemble into virus-like particles (VLPs) and prevent infection by eliciting neutralizing antibodies. Adding the other frequently identified oncogenic types 31, 33, 45, 52, and 58 to a vaccine would increase the coverage against HPV-induced cancers to approximately 90%. We describe the generation and characterization of panels of monoclonal antibodies to these five additional oncogenic HPV types, and the selection of antibody pairs that were high affinity and type specific and recognized conformation-dependent neutralizing epitopes. Such characteristics make these antibodies useful tools for monitoring the production and potency of a prototype vaccine as well as monitoring vaccine-induced immune responses in the clinic.

scholarly journals Potential Use of Salivary Markers for Longitudinal Monitoring of Inflammatory Immune Responses to Vaccination

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Pei Wen Lim ◽  
Johan Garssen ◽  
Elena Sandalova
Keyword(s):  
Immune Responses ◽  
Time Course ◽  
Dynamic Range ◽  
Inflammatory Responses ◽  
Literature Survey ◽  
Inflammatory Biomarkers ◽  
Immune Reactivity ◽  
Kinetics Of ◽  
Potential Use ◽  
Full Dynamic Range

Vaccination, designed to trigger a protective immune response against infection, is a trigger for mild inflammatory responses. Vaccination studies can address the question of inflammation initiation, levels, and resolution as well as its regulation for respective studied pathogens. Such studies largely based on analyzing the blood components including specific antibodies and cytokines were usually constrained by number of participants and volume of collected blood sample. Hence, blood-based studies may not be able to cover the full dynamic range of inflammation responses induced by vaccination. In this review, the potential of using saliva in addition to blood for studying the kinetics of inflammatory response studies was assessed. Saliva sampling is noninvasive and has a great potential to be used for studies aimed at analysing the magnitude, time course, and variance in immune responses, including inflammation after vaccination. Based on a literature survey of inflammatory biomarkers that can be determined in saliva and an analysis of how these biomarkers could help to understand the mechanisms and dynamics of immune reactivity and inflammation, we propose that the saliva-based approach might have potential to add substantial value to clinical studies, particularly in vulnerable populations such as infants, toddlers, and ill individuals.

Preparation and Immunological Traits of Monoclonal Antibody against Sarafloxacin

2012 ◽  
Vol 459 ◽  
pp. 54-57
Author(s):  
Guo Ying Fan ◽  
Jin Qing Jiang
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Cell Fusion ◽  
Light Chain ◽  
Dynamic Range ◽  
Fusion Technology ◽  
Alternative Method ◽  
Ic50 Values ◽  
The Stability ◽  
Igg1 Isotype

Through cell fusion technology, five hybridoma lines of sarafloxacin (SAR), named S1-B2, S2-C6, S2-E7, S3-C5, and S3-E5, were identified and their corresponding mAbs were of the IgG1 isotype with a k light chain. The Kaffs of all mAbs were between 2.8 and 4.6×109 L/mol. The titers and IC50 values of purified ascite fluids were in the range of 0.512–2.56×106 and 0.32–0.48 ng/mL, respectively. The performances of S1-B2 and S2-C6 were more consistent in the stability experiments. Based on the S1-B2 hybridoma, an icELISA method was developed. The dynamic range was from 0.004 to 18 ng/mL, with a detection limit for the assay and IC50 values of 0.002 and 0.32 ng/mL, respectively. Therefore, the establishment of these hybridomas may provide an alternative method for the detection of SAR residues in food-original animals.

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