scholarly journals Nanostructured recombinant protein particles raise specific antibodies against the nodavirus NNV coat protein in sole

2020 ◽  
Vol 99 ◽  
pp. 578-586 ◽  
Author(s):  
Rosemary Thwaite ◽  
Concepción Berbel ◽  
Manuel Aparicio ◽  
Debora Torrealba ◽  
Mireia Pesarrodona ◽  
...  
Keyword(s):  
Coat Protein ◽  
Recombinant Protein ◽  
Specific Antibodies ◽  
Protein Particles

Selective Release of Recombinant Protein Particles (VLPs) from Yeast Using a Pure Lytic Glucanase Enzyme

1993 ◽  
Vol 11 (2) ◽  
pp. 214-217 ◽  
Author(s):  
J. A. Asenjo ◽  
A. M. Ventom ◽  
R. -B. Huang ◽  
B. A. Andrews
Keyword(s):  
Recombinant Protein ◽  
Protein Particles

scholarly journals Reproductive and Behavioral Evaluation of a New Immunocastration Dog Vaccine

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 226
Author(s):  
Daniela Siel ◽  
María José Ubilla ◽  
Sonia Vidal ◽  
Alexandra Loaiza ◽  
John Quiroga ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Recombinant Protein ◽  
Control Group ◽  
Low Molecular Weight ◽  
Specific Antibodies ◽  
Controlled Conditions ◽  
Low Molecular Weight Chitosan ◽  
And Control ◽  
Complementary Strategy ◽  
Positive Effect

Canine immunocastration development has been of interest for many years as a complementary strategy to surgical castration. The purpose of this paper was to verify the effect of a recombinant vaccine for dog immunocastration. Two tests were done, one under controlled conditions and a second under field conditions. Animals were injected with 1 mL of 500 µg GnRXG/Q recombinant protein; 500 µg of low molecular weight chitosan as adjuvant; 1 mL NaCl 0.9% q.s. In the first trial, eight Beagle male dogs between the ages of 1 and 3 comprised the sample, randomly divided into two groups: vaccinated group (n = 7) and control group (n = 2). The second trial had 32 dogs with owners. In the first controlled conditions trial, the vaccine produced specific antibodies that remained until the end of the trial (day 270), inducing reduced testosterone and spermiogram changes in the immunized animals. In a second trial, on the field, specific immunity was induced, which remained high up to day 150. The vaccine also reduced sexual agonistic and marking behaviors. This new vaccine proved to be safe, immunogenic, capable of reducing gonadal functionality, and had a positive effect on inducing reduced sexual, agonistic, and marking behavior of the animals.

scholarly journals Antibodies Reactive with the N-Terminal Domain ofPlasmodium falciparum Serine Repeat Antigen Inhibit Cell Proliferation by Agglutinating Merozoites and Schizonts

1999 ◽  
Vol 67 (4) ◽  
pp. 1821-1827 ◽  
Author(s):  
Xin-Li Pang ◽  
Toshihide Mitamura ◽  
Toshihiro Horii
Keyword(s):  
Recombinant Protein ◽  
Parasitophorous Vacuole ◽  
Parasite Growth ◽  
Specific Antibodies ◽  
Terminal Domain ◽  
Rbc Membrane ◽  
Ofplasmodium Falciparum ◽  
Vaccine Candidate Antigen ◽  
Specific Igg

ABSTRACT The serine repeat antigen (SERA) is a vaccine candidate antigen ofPlasmodium falciparum. Immunization of mice withEscherichia coli-produced recombinant protein of the SERA N-terminal domain (SE47′) induced an antiserum that was inhibitory to parasite growth in vitro. Affinity-purified mouse antibodies specific to the recombinant protein inhibited parasite growth between the schizont and ring stages but not between the ring and schizont stages. When Percoll-purified schizonts were cultured with the affinity-purified SE47′-specific antibodies, schizonts and merozoites were agglutinated. Indirect-immunofluorescence assays with unfixed parasite cells showed that SE47′-specific immunoglobulin G (IgG) bound to SERA molecules on rupturing schizonts and merozoites but the IgG did not react with the schizont-infected erythrocytes (RBC). Furthermore, double-fluorescence staining against SE47′-specific IgG and anti-human RBC membrane IgG showed that the RBC membrane disappeared from SE47′-specific-IgG-bound schizonts after cultivation. These observations suggest that the SE47′-specific antibodies inhibit parasite growth by cross-linking SERA molecules that are associated with merozoites in rupturing schizonts with partly broken RBC and parasitophorous vacuole membranes, blocking merozoite release.

Immunoaffinity purification of bluetongue virus group specific antibody using recombinant protein adsorbed to polystyrene wells

2017 ◽  
Author(s):  
Karam Chand ◽  
Sanchay K. Biswas ◽  
Bimalendu Mondal ◽  
Awadh B. Pandey
Keyword(s):  
Recombinant Protein ◽  
Specific Antibody ◽  
Bluetongue Virus ◽  
Affinity Purification ◽  
Disease Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Immunoaffinity Purification ◽  
Purification Technique ◽  
Hybridoma Technology

Most of the common viral diseases can be diagnosed by serological assays and these assays mostly depend on the purity and quality of antibody used. Such specific antibodies can be generated by hybridoma technology. Alternatively, the virus-specific antibodies can be purified from polyclonal serum by immunoaffinity purification technique. Based on this immune affinity purification technique we have purified group-specific antibody against Bluetongue virus (BTV) using recombinant protein VP7 of BTV bound to polystyrene wells. Elution buffer consisting of 100 mM Glycine-HCl, pH 3.0 was found optimum for dissociation of the antibody from recombinant antigen and also to maintain the integrity of antigen. The reactivity of eluted antibody was tested in enzyme-linked immunosorbent assay (ELISA). The purified antibody will be useful in other serological assays like ELISA, fluorescent assay, and agar gel immunodiffusion (AGID) for Bluetongue disease diagnosis

scholarly journals Surface Engineering of Recombinant RNA Coliphage Qβ to Display gp41 MembraneProximal External-Region Epitopes from HIV-1

2017 ◽  
Vol 2 (1) ◽  
Keyword(s):  
Immune System ◽  
Coat Protein ◽  
Antibody Responses ◽  
Igg Antibody ◽  
External Region ◽  
Specific Antibodies ◽  
Infected People ◽  
Specific Igg ◽  
Minor Coat Protein ◽  
Hiv 1

Introduction: The membrane proximal external region (MPER) of HIV-1 envelope glycoprotein-41 (gp41) is targeted by several broadly neutralizing antibodies whose conserved linear epitopes are promising targets for vaccine design. However, a formidable challenge has remained the difficulty to design and deliver MPER based immunogens for the efficient induction of such broadly neutralizing HIV-1 specific antibodies (bnAb). This is mainly because the linear bnAb MPER epitopes are poorly accessible to the immune system. The overall objective of this study therefore was the development of a novel RNA Qβ phage display system not only for monitoring anti-MPER specific antibody responses but equally as potential immunogens in future HIV-1 vaccine designs. Method: To overcome the challenge of effective presentation of MPER to the immune system we have selectively engineered the surface of the RNA coliphage Qβ to expose all MPER bnAb epitopes. Briefly, DNA representing a 50 amino acids consensus region within the HIV-1 gp41 MPER was fused in frame with the minor coat protein A1of the Qβ phage. Three variant MPER expression cassettes were obtained with the MPER cDNA in frame with the minor coat protein A1 gene, including pQβMPER, pQβMPERHis and pQβMPERN. The expression cassettes were used for the production of QβMPER recombinant phages after transformation of E. coli HB101 strain. Antigencity of the phages was assessed with plasma from long standing anti-retroviral naïve HIV-1 infected people from the CIRCB AFRODEC cohort while immunogenicity studies were done in female Balb/c mice. Results: The initial titers of all recombinant phages including QβMPER, QβMPERHis and QβMPERN were 104 plaque forming units/ml (pfu/ml). This was significantly lower (P<0.001) relative to the 108 pfu/ml of wild type phage, but was scaled up to 1014pfu/ml. The fusion of MPER and Qβ genes was confirmed by RT-PCR followed by gel electrophoresis and sequencing. Specific recognition of some reported bnAb epitopes within MPER were confirmed in ELISA using the three recombinant QβMPER phages together with an MPER restrictive peptide as antigens and the bnAb 4E10, Z13e1, 2F5 and 10E8 as antibodies. Next the prevalence of MPER-specific antibodies was determined in plasma from long standing antiretroviral naïve HIV-1 infected participants of the CIRCB AFRODEC cohort. The greater majority (84%) of participants’ plasma showed MPER peptide specific reactivity with anti-MPER specific IgG antibody titers ranging from 200 to 409600 comparative to background IgG antibody titer with the Qβ phage alone as antigen or plasma from seronegative participants. In immunogenicity studies in Balb/c mice the recombinant phages QβMPERN and QβMPERHis induced significantly high Anti-MPER-specific IgG antibody responses (P<0.04) in at least 60 % of mice following three inoculations of each recombinant phage. Conclusion: Thus, these novel recombinantQβMPER phages can be used to monitor MPER-specific immune responses in HIV-1 exposed or infected people. In addition the recombinant QβMPER phages could be used as immunogens either alone as demonstrated here in mice or in combination with other strategies for the induction of MPER specific immunity against HIV-1.

scholarly journals Тransplastomic tobacco plants producing the hydrophilic domain of the sheep pox virus coat protein L1R

2020 ◽  
Vol 24 (8) ◽  
pp. 905-912
Author(s):  
D. K. Beisenov ◽  
G. E. Stanbekova ◽  
B. K. Iskakov
Keyword(s):  
Coat Protein ◽  
Recombinant Protein ◽  
Blot Analysis ◽  
Neutralizing Antibodies ◽  
Plastid Dna ◽  
Plant Genome ◽  
Total Soluble Protein ◽  
Economic Losses ◽  
Pcr Analysis ◽  
Future Research

Sheep pox has a wide geographical range of distribution and poses a threat to sheep breeding worldwide, as the disease is highly contagious and is accompanied by large economic losses. Vaccines based on live attenuated virus strains are currently being used for prevention of this disease. Such vaccines are effective, but potentially dangerous because of the possible virus reversion to a pathogenic state. The development of safe recombinant subunit vaccines against sheep pox is very relevant. The high ploidy level of the plant chloroplasts makes it possible to obtain large quantities of foreign proteins. The purpose of this study was to create transplastomic Nicotiana tabacum plants producing one of the candidate vaccine proteins of sheep pox virus L1R. A vector containing a deletion variant of the SPPV_56 gene, which encodes the N-terminal hydrophilic part of the viral coat protein L1R, was constructed to transform tobacco plastids. It provides integration of the transgene into the trnG/trnfM region of the chloroplast tobacco genome by homologous recombination. Spectinomycin-resistant tobacco lines were obtained by biolistic gun-mediated genetic transformation. PCR analysis in the presence of gene-specific primers confirmed integration of the transgene into the plant genome. Subsequent Northern and Western blot analysis showed the gene expression at the transcriptional and translational levels. The recombinant protein yields reached up to 0.9 % of total soluble protein. The transplastomic plants displayed a growth retardation and pale green leaf color compared to the wild type, but they developed normally and produced seeds. Southern blot analysis showed heteroplasmy of the plastids in the obtained plants due to recombination events between native and introduced regulatory plastid DNA elements. The recombinant protein from plant tissue was purified using metal affinity chromatography. Future research will be focused on determining the potential of the chloroplast-produced protein to induce neutralizing antibodies against SPPV strains.

Sign in / Sign up

Export Citation Format

Share Document