Polyclonal Antibodies Elicited via Immunization with a Ru(bpy)32+-Methyl Viologen Conjugate: Is a Polyclonal Antibody Immune Response Always Heterogeneous?

1995 ◽  
Vol 117 (9) ◽  
pp. 2673-2674 ◽  
Author(s):  
Kevin Shreder ◽  
Anthony Harriman ◽  
Brent L. Iverson
Keyword(s):  
Immune Response ◽  
Polyclonal Antibody ◽  
Methyl Viologen ◽  
Polyclonal Antibodies

scholarly journals Polyclonal antibody catalytic variability

1998 ◽  
Vol 332 (1) ◽  
pp. 127-134 ◽  
Author(s):  
David B. STEPHENS ◽  
Richard E. THOMAS ◽  
John F. STANTON ◽  
Brent L. IVERSON
Keyword(s):  
Immune Response ◽  
Catalytic Activity ◽  
Polyclonal Antibody ◽  
Specific Binding ◽  
Polyclonal Antibodies ◽  
Catalytic Activities ◽  
Animal System ◽  
Antibody Catalysis ◽  
Variability Study ◽  
Accurate Indicator

We have performed a systematic variability study of polyclonal antibody catalysis by using five rabbits immunized with the same hapten. Important results from this work are the following. (1) Similarities were observed in the catalytic polyclonal antibodies derived from all five rabbits. Four of the five rabbits produced polyclonal samples that were nearly the same in terms of catalytic activity, whereas the fifth rabbit, designated as rabbit 2, displayed a somewhat higher level of catalytic activity. The catalytic activities (as kcat/kuncat) of these polyclonal samples were similar to that from the best murine monoclonal antibody that had been previously elicited by the same hapten. (2) Titre was not an accurate indicator of polyclonal antibody catalytic activity. (3) A mathematical analysis to describe a distribution of Michaelis–Menten catalysts was performed to help interpret our results. (4) Kinetic analysis indicated that the binding parameters of the different samples were remarkably homogeneous, because one or two components were all that were required to fit the on-rate and off-rate data satisfactorily. Interestingly, the most active catalytic polyclonal sample, that from rabbit 2, displayed the slowest off-rate (so slow it could not be measured) and thus the highest overall affinity. (5) Catalytic analysis of eluted fractions of antibody from a substrate column indicated that each polyclonal sample was also relatively homogeneous in terms of catalytic parameters. The main conclusion of our study is that for this hapten–animal system, the overall catalytic immune response is relatively consistent at two levels. Consistent catalytic activity was observed between the polyclonal samples elicited in the different animals, and the elicited hapten-specific polyclonal antibodies were relatively homogeneous in terms of binding and catalytic parameters within each immunized animal. The observed similarities of the catalytic activity in the different animals is surprising, because the immune response is based on specific binding of antibodies to hapten. There is no known selective pressure to maintain consistent levels of catalytic activity. Our results can therefore be interpreted as providing evidence that for this hapten there is a fixed relationship between hapten structure and catalytic activity and/or consistent genetic factors that dominate the catalytic immune response.

Evaluation of germ plasm for resistant to Soybean mosaic virus (SMV)

2020 ◽  
Vol 21 (1) ◽  
pp. 6-9
Author(s):  
Wuye Ria Andayanie
Keyword(s):  
Abiotic Stress ◽  
Environmental Factors ◽  
Mosaic Virus ◽  
Polyclonal Antibody ◽  
Symptom Severity ◽  
Germ Plasm ◽  
Polyclonal Antibodies ◽  
Soybean Mosaic Virus ◽  
High Yield ◽  
High Yields

Soybean superior varieties with high yields and are resistant to abiotic stress have been largely released, although some varieties grown in the field are not resistant to SMV. In addition, the opportunity to obtain lines of hope as prospective varieties with high yield and resistance to SMV is very small. The method for evaluating soybean germplasm is based on serological observations of 98 accessions of leaf samples from SMV inoculation with T isolate. The evaluation results of 98 accessions based on visual observations showed 31 genotypes reacting very resistant or healthy to mild resistant category to SMV T isolate  with a percentage of symptom severity of 0 −30 %. Among 31 genotypes there are 2 genotypes (PI 200485; M8Grb 44; Mlg 3288) with the category of visually very resistant and resistant, respectively and  Mlg 3288  with the category of mild resistant.  They have a good agronomic appearance with a weight of 100 seeds (˃10 g) and react negatively with polyclonal antibodies to SMV, except Mlg 3288 reaction is not consistent, despite the weight of 100 seeds (˃ 10 g). Leaf samples from 98 accessions revealed various symptoms of SMV infection in the field. This diversity of symptoms is caused by susceptibility to accession, when infection occurs, and environmental factors. Keywords—: soybean; genotipe; Soybean mosaic virus (SMV); disease severity; polyclonal  antibody

Immunofluorescence Assay Using Monoclonal and Polyclonal Antibodies for Detection of Staphylococcal Enterotoxins A in Milk

2019 ◽  
Vol 13 (1) ◽  
pp. 137-145 ◽  
Author(s):  
Tzonka Godjevargova ◽  
Zlatina Becheva ◽  
Yavor Ivanov ◽  
Andrey Tchorbanov
Keyword(s):  
Monoclonal Antibody ◽  
Magnetic Nanoparticles ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Food Poisoning ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin A ◽  
Fluorescence Immunoassay ◽  
Magnetic Separator ◽  
Milk Samples

Objectives: Staphylococcus aureus is a Gram-positive microorganism. S. aureus can grow in various foods and cause food poisoning by secreting enterotoxins. The most common enterotoxins involved in food poisoning are staphylococcal enterotoxin A and staphylococcal enterotoxin B, but Staphylococcal Enterotoxin A (SEA) is predominant. The main types of food contaminated with SEs are meat and meat products, poultry and eggs, milk and dairy products. The aim of this study was to develop a rapid and sensitive fluorescence immunoassay for detection of staphylococcal enterotoxin A in milk. Methods: Monoclonal and polyclonal antibodies for SEA were produced and characterized. Competitive fluorescence immunoassay based on Magnetic Nanoparticles (MNPs) was performed and optimized. MNPs were used as a solid carrier of the antibodies. The first step of the assay was immunoreaction between the immobilized antibody onto MNPs and SEA in milk sample. Then the fluorescein-SEA conjugate was added to the sample. Thus, competitive immunoreaction between MNP-mAb/MNP-pAb with SEA and SEA-FITC was performed. These immuno-complexes were separated by a magnetic separator and the obtained supernatants were analyzed. The fluorescent signal from the excess of conjugated SEA was proportional to the SEA contained in the milk. The assay duration was only 30 min. Results: The fluorescence immunoassays performed with polyclonal antibody had linear ranges from 5 pg/mL to 100 ng/mL SEA in a buffer, and from 50 pg/mL to 50 ng/mL SEA in spiked milk samples. While the same assays performed with monoclonal antibody had linear ranges from 1 pg/mL to 20 ng/mL SEA in buffer, and from 10 pg/mL to 10 ng/mL SEA in spiked milk samples. The detection limits of the developed immunoassays performed in milk were: 48 pg/mL with polyclonal antibody and 9 pg/mL with monoclonal antibody. Conclusion: A rapid and sensitive fluorescence immunoassay based on magnetic nanoparticles with a polyclonal and monoclonal antibody for determination of staphylococcal enterotoxin A in milk was developed.

scholarly journals Differential expression of TgMIC1 in isolates of Chinese 1 Toxoplasma with different virulence

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yang Wang ◽  
Chengjian Han ◽  
Rongsheng zhou ◽  
Jinjin Zhu ◽  
Famin Zhang ◽  
...  
Keyword(s):  
Differential Expression ◽  
Rna Sequencing ◽  
Polyclonal Antibody ◽  
Protein Level ◽  
Polyclonal Antibodies ◽  
Mrna Levels ◽  
Quantitative Real Time Pcr ◽  
Sequencing Data ◽  
Predominant Genotype ◽  
High Virulence

Abstract Background The predominant genotype of Toxoplasma in China is the Chinese 1 (ToxoDB#9) lineage. TgCtwh3 and TgCtwh6 are two representative strains of Chinese 1, exhibiting high and low virulence to mice, respectively. Little is known regarding the virulence mechanism of this non-classical genotype. Our previous RNA sequencing data revealed differential mRNA levels of TgMIC1 in TgCtwh3 and TgCtwh6. We aim to further confirm the differential expression of TgMIC1 and its significance in this atypical genotype. Methods Quantitative real-time PCR was used to verify the RNA sequencing data; then, polyclonal antibodies against TgMIC1 were prepared and identified. Moreover, the invasion and proliferation of the parasite in HFF cells were observed after treatment with TgMIC1 polyclonal antibody or not. Results The data showed that the protein level of TgMIC1 was significantly higher in high-virulence strain TgCtwh3 than in low-virulence strain TgCtwh6 and that the invasion and proliferation of TgCtwh3 were inhibited by TgMIC1 polyclonal antibody. Conclusion Differential expression of TgMIC1 in TgCtwh3 and TgCtwh6 may explain, at least partly, the virulence mechanism of this atypical genotype.

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 THE VALUE OF THE IMMUNE RESPONSE IN PATIENTS WITH EBV INFECTIOUS MONONUCLEOSIS IN PREDICTING THE COURSE AND EFFICACY OF ANTIVIRAL AND IMMUNO IMMUNOCORRECTING THERAPY

2013 ◽  
Vol 18 (1) ◽  
pp. 7-14
Author(s):  
T. A. Svintsova ◽  
D. M. Sobchak ◽  
O. V. Korochkina ◽  
G. A Kravchenko ◽  
V. V Novikov
Keyword(s):  
Immune Response ◽  
Infectious Mononucleosis ◽  
Epstein Barr Virus ◽  
Mononuclear Cells ◽  
Polyclonal Antibodies ◽  
Severity Of Illness ◽  
Control Group ◽  
Differentiation Antigens ◽  
Barr Virus ◽  
Epstein Barr

The indices of immune response were studied in 68 patients with infectious mononucleosis caused by the Epstein-Barr virus (35 males, 33 females) aged 18 to 30 years. Materials and methods. The content of soluble forms of differentiation antigens (sCD95, sCD18, sCD50, sHLAI, sCD54) has been studied with enzyme immunoassay using monoclonal antibodies Mab IC0-20 and polyclonal antibodies to the antigens of the mononuclear cells of the peripheral blood. The control group included 60 healthy volunteers matched for age and sex with the main group. The aim of this study is the assessment of the content of soluble forms of differentiation antigens in patients with infectious mononucleosis caused by Epstein-Barr virus, depending on gender, age, severity of illness, comorbidities, laboratory values, the presence of viral DNA, as well as a demonstration of their value in predicting the course and outcome of the disease and the efficacy of antiviral and immunocorrecting therapy. In patients with negative results of DNA indication of EBV a significant increase in the content of soluble forms of differentiation antigens characterizing the adhesion of leukocytes (sCD18), the activity of T-lymphocytes (sCD50), the recognition of foreign antigens (sHLAI) in the blood in comparison with patients with a positive DNA indication of EBV was determined. Conclusion. According to the results of this performed work the criterion for an adequate immune response in patients with infectious mononucleosis caused by the Epstein-Barr virus was found to be the increase of the content of soluble forms of differentiation antigens (sCD95, sCD18, sCD50 sHLAI, sCD54). In patients with exanthema, tonsillar syndrome, leukocytosis, elevation of transaminases and the presence of antibodies to capsid antigen (a/VCAIgM) the content of soluble forms of differentiation antigens (sCD95, sCD18, sCD50 sHLAI, sCD54), was higher than in patients without such symptoms. In the treatment with cycloferon in patients with cyclic course of EBV infectious mononucleosis the content of sHLAI and sCD54 at 2nd-4th weeks of treatment increased by 1.5-2 times compared with the corresponding values before treatment. In patients with reactivation of the disease monotonically low indices of all studied soluble forms of differentiation antigens persisted over the 4 weeks during patients following up. In patients with infectious mononucleosis caused by Epstein-Barr virus, the dynamics of sHLAI and sCD54 after 2-4 weeks of treatment serves as secondary efficacy endpoint of antiviral, immunomodulatory therapy and the formation of the cyclic course of the disease.

scholarly journals The Ability of Zika virus Intravenous Immunoglobulin to Protect From or Enhance Zika Virus Disease

2021 ◽  
Vol 12 ◽  
Author(s):  
Amelia K. Pinto ◽  
Mariah Hassert ◽  
Xiaobing Han ◽  
Douglas Barker ◽  
Trevor Carnelley ◽  
...  
Keyword(s):  
Virus Infection ◽  
Polyclonal Antibody ◽  
Zika Virus ◽  
Virus Disease ◽  
Polyclonal Antibodies ◽  
Antibody Therapeutics ◽  
The World ◽  
Flavivirus Infection

The closely related flaviviruses, dengue and Zika, cause significant human disease throughout the world. While cross-reactive antibodies have been demonstrated to have the capacity to potentiate disease or mediate protection during flavivirus infection, the mechanisms responsible for this dichotomy are still poorly understood. To understand how the human polyclonal antibody response can protect against, and potentiate the disease in the context of dengue and Zika virus infection we used intravenous hyperimmunoglobulin (IVIG) preparations in a mouse model of the disease. Three IVIGs (ZIKV-IG, Control-Ig and Gamunex®) were evaluated for their ability to neutralize and/or enhance Zika, dengue 2 and 3 viruses in vitro. The balance between virus neutralization and enhancement provided by the in vitro neutralization data was used to predict the IVIG concentrations which could protect or enhance Zika, and dengue 2 disease in vivo. Using this approach, we were able to define the unique in vivo dynamics of complex polyclonal antibodies, allowing for both enhancement and protection from flavivirus infection. Our results provide a novel understanding of how polyclonal antibodies interact with viruses with implications for the use of polyclonal antibody therapeutics and the development and evaluation of the next generation flavivirus vaccines.

scholarly journals IMMUNODIAGNOSIS INFEKSI Aeromonas hydrophila PADA IKAN

2018 ◽  
Vol 36 (1) ◽  
pp. 80
Author(s):  
Yuli Purwandari Kristianingrum ◽  
Sitarina Widyarini ◽  
Kurniasih Kurniasih ◽  
Bambang Sutrisno ◽  
Charles Rangga Tabbu ◽  
...  
Keyword(s):  
Blood Serum ◽  
Polyclonal Antibody ◽  
Aeromonas Hydrophila ◽  
Intraperitoneal Injection ◽  
Staining Method ◽  
Polyclonal Antibodies ◽  
Ulcer Disease ◽  
Tube Test ◽  
Immunohistochemistry Staining ◽  
Motile Aeromonas Septicaemia

Aeromonas hydrophila causes a disease that often infects fish and is known as Motile Aeromonas Septicaemia (MAS), Hemorrhagi Septisemia, Ulcer disease or Red-Sore disease. The   aims of this study were to develop polyclonal antibody of  Aeromonas hydrophila in the rabbits to   confirm the diagnosis of Aeromonas hydrophila  in the fish by immunohistochemistry staining method. Preparation of polyclonal antibodies was performed on the rabbits used to Aeromonas hydrophila bacteria that have been tested biochemically by intravenous and intraperitoneal injection. Doses of Aeromonas hydrophila  bacteria were 109 CPU/ml  of 0.5 ml at first injection, 1 ml at second injection, 2 ml at thirth injection and 3 ml at fourth injection. Blood serum collection was performed at week 5 after injection from  an  ear and intracardial vein. The result of antibody titer was 28 = 1024 which measured by   tube test. Furthermore, polyclonal   antibody was used to immunohistochemistry  staining with 400x dilution. The results of the staining showed that an immunopositive reaction in the liver, skin,lien,  gill, kidney, and heart of fish to Aeromonas hydrophila antibody. The research conclution was polyclonal antibody from rabbit can be used to accurately confirm the diagnosis of Aeromonas hydrophila  based on antigen and antibody reaction. 

scholarly journals Subclass-restricted IgG polyclonal antibody production in mice injected with lipid A-rich lipopolysaccharides.

1981 ◽  
Vol 153 (2) ◽  
pp. 324-338 ◽  
Author(s):  
S Izui ◽  
R A Eisenberg ◽  
F J Dixon
Keyword(s):  
Nude Mice ◽  
Polyclonal Antibody ◽  
Lipid A ◽  
Polyclonal Antibodies ◽  
Serum Levels ◽  
Igg Subclass ◽  
Igg Antibodies ◽  
Athymic Nude Mice ◽  
Bacterial Lipopolysaccharides ◽  
Autoantibody Formation

The effects of five distinct bacterial lipopolysaccharides (LPS) on the induction of polyclonal IgM and IgG antibodies, including polyclonal autoantibody formation, were investigated in several strains of mice. Injections of most LPS preparations that contained polysaccharide transiently induced only IgM polyclonal antibodies. However, LPS from Salmonella minnesota R595 (R595 LPS), which had a particularly high content of lipid A but lacked O-antigen polysaccharide, induced a markedly prolonged IgM and IgG polyclonal antibody response in mice, including athymic nude mice, but not in LPS-unresponsive C3H/HeJ mice. Polyclonal IgM and IgG production peaked in sera on day 8 and day 15, respectively, and remained higher than control values 2 mo after the injection. The IgG induced by R595 LPS was strictly restricted to IgG2b and Igg3 subclasses in normal mice. In contrast, in athymic nude mice which have normally lower levels of IgG1 and IgG2a than normal mice, R595 LPS stimulated the production of all the IgG subclasses and reconstituted serum levels of IgG1 and IgG2a up to, but not higher than, control values of normal mice. These findings suggest that different mechanisms regulate production of each IgG subclass after stimulation with LPS.

scholarly journals The third member of the Tetrahymena CCT subunit gene family, TpCCTα, encodes a component of the hetero-oligomeric chaperonin complex

1997 ◽  
Vol 326 (1) ◽  
pp. 21-29 ◽  
Author(s):  
Helena SOARES ◽  
Luisa CYRNE ◽  
Cristina CASALOU ◽  
Bruno EHMANN ◽  
Claudina RODRIGUES-POUSADA
Keyword(s):  
Gene Expression ◽  
Gene Family ◽  
Polyclonal Antibody ◽  
Similar Pattern ◽  
Synthetic Peptides ◽  
Polyclonal Antibodies ◽  
Recombinant Yeast ◽  
Subunit Gene ◽  
The Third ◽  
Tubulin Genes

The sequence of a third member of the Tetrahymenapyriformis chaperonin CCT (‘chaperonin containing TCP1’) subunit gene family is presented. This gene, designated TpCCTα, is the orthologue of the mouse chaperonin gene TCP1/CCTα. To characterize the CCT complex in this ciliate, we have produced polyclonal antibodies against synthetic peptides based on C-terminal sequences deduced from the primary sequences of the TpCCTα, TpCCTγ and TpCCTη subunits. We have also used polyclonal antibodies produced against recombinant yeast CCTα and CCTβ subunits. Using these antibodies, we show that Tetrahymena cells contain a hetero-oligomeric CCT chaperonin comprising at least seven distinct subunits. Three of these were assigned to specific TpCCT genes, whereas a fourth was recognized by the polyclonal antibody against yeast CCTβ, suggesting that this gene is also present in the ciliate. The CCT complex also contains other unidentified proteins that were recognized by the polyclonal antibody UM-1, raised against the putative ATP binding domain of the chaperonin proteins. TpCCTα gene expression was shown in exponentially growing cells and cells regenerating their cilia for different periods to have a similar pattern to the previously identified genes TpCCTγ and TpCCTη, and also to tubulin genes.

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