جستجوی مقالات مرتبط با کلیدواژه "recombinant antigen" در نشریات گروه "پزشکی"

Echinococcosis or hydatid disease is one of the most important zoonosis across the world.

The recombinant P29 protein is considered as a suitable candidate for evaluating diagnostic ELISA to reach a more accurate conclusion.

In this study the P29 sequence comprised with somewhat similar sequences registered in Gene Bank and showed 100% identity with P29 sequences derived from Echinococcus granolosus (E. granulosus) (XP_024351425.1), E. multilocularis (AHA85396.1), Endophilin B1 protein of E. granulosus (CDS21096.1). The primary and secondary structure of the peptide was analyzed due to its characterization of peptide motifs to be expressed on MHC Class HLA allels.

The recombinant P29 derived peptides can be produced and analyzed due to be determined as vaccine design candidate against Hydatidosis.

Among all types of malignant diseases, breast cancer has worldwide importance because of the high mortality rate in women aged fewer than 50 in the developing countries. Identification of immunogenic antigens and the generation of specific antibodies against cancer cells are the most successful strategies for early detection and effective treatment of breast cancer. In the current study, a chimeric protein consisting of two specific surface antigens, MUC1 and HER2, were used for the production of chitosan nanoparticles and evaluated as a vaccine candidate. The pET-28a expression vector harboring the HER2-MUC1 gene was constructed. Expression of the protein in E. coli BL21 (DE3) was induced using IPTG. The recombinant HER2-MUC1 (HM) protein was purified using a Ni-NTA column and confirmed by western blotting. Chitosan nanoparticles containing the target protein were prepared and the lymphocytes viability was evaluated using MTT assay. The expression of the recombinant protein with molecular weight of 40 kDa was confirmed using SDS-PAGE and Western blotting. The electric charge and the size of the nanoparticles were determined and verified by a Zeta Sizer device. The evaluation of IgG and IgA titration suggested that inducing humoral and mucosal immune responses by administering nanoparticles containing the chimeric protein. Analysis of cell-mediated immunity showed that the chimeric HM protein could induce specific splenocyte proliferation in immunized mice. It seems that HM nanoparticles can be utilized as a vaccine candidate for inducing the cellular and humoral immune response against breast cancer.

Breast cancer is one of the most common cancers in the world and is on the increase. MUC1 and HER2 as tumor-associated antigens (TAAs) are abnormally expressed to some extent in 75–80% of breast cancers. In our present research, a novel chimeric MUC1-HER2 (HM) protein was designed and used to study whether an immune response can be generated against these TAAs. In vitro analysis of the HER2-MUC1 construct confirmed the co-expression of MUC1 and HER2.
BALB/c mice were immunized with this novel chimeric protein. The humoral immune response was assessed by enzyme-linked immunosorbent assay (ELISA). Then, BALB/c mice were injected subcutaneously 2×105 4T1-MUC1-HER2 tumor cells. Subsequently, tumor size and tumor necrosis measurements, MTT, cytokines assay and survival test were performed.
The results implied a critical role of HER2 and MUC1 antibodies in vaccination against breast cancer. This engineered protein can be a good vaccine to stop breast cancer.
The results implied a critical role of HER2 and MUC1 antibodies in vaccination against breast cancer. This engineered protein can be a good vaccine to stop breast cancer.

Toxoplasma gondii (T. gondii) is distributed worldwide and infects most species. The serious incidence and severe or fatal injury caused by T. gondii infection clearly indicates the necessity for the event of a vaccine. The current study goals were to evaluate serological applications of Toxoplasma gondiirhoptry protein 1 (ROP1) antigen.

We created a polymer vaccine by using the eukaryotic plasmid, pROP1. Purification by one-step metal affinity chromatography allowed recovery of milligram amounts of purified recombinant proteins per liter of culture. The quality of this matter for diagnosing of human infections was provided and tested on 77 serum samples which were obtained during routine diagnostic tests. A panel of 20 serum samples from patients with acute toxoplasmosis was compared to a panel of 35 serum samples from individuals with chronic toxoplasmosis.

Results of the study indicated that antibodies detected from patients with acute and chronic infections were 96% and 17%, respectively, by using of pROP1 recombinant antigen.

According to the present study an immunoglobulin G antibody against ROP1 antigen is made throughout the acute stage of toxoplasmosis infection, but not in the chronic phase of toxoplasmosis.

Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory.
The current study aimed at sub-cloning a gene, encoding the B8/1 subunit of antigen B (AgB) from Echinococcus granulosus, using gene optimization for the immunodiagnosis of human CE.
The coding sequence for AgB8/1 subunit of Echinococcus granulosus
was selected from GenBank and was gene-optimized. The sequence was synthesized
and inserted into pGEX-4T-1 vector. Purification was performed with GST tag affinity
column. Diagnostic performance of the produced recombinant antigen, native antigen B
and a commercial ELISA kit were further evaluated in an ELISA system, using a panel
of sera from CE patients and controls.
SDS-PAGE demonstrated that the protein of interest had a high expression level and purity after GST tag affinity purification. Western blotting verified the immunoreactivity of the produced recombinant antigen with the sera of CE patients. In an ELISA system, the sensitivity and specificity (for human CE diagnosis) of the recombinant antigen, native antigen B and commercial kit were respectively 93% and 92%, 87% and 90% and 97% and 95%.
The produced recombinant antigen showed a high diagnostic value which can be recommended for serodiagnosis of CE in Iran and other CE-endemic areas. Utilizing the combination of other subunits of AgB8 would improve the performance value of the introduced ELISA system.

Dense granules are immunodominant proteins for the standardiza­tion of immunodiagnostic procedures to detect neosporosis. In the presented study different fragment of a dense-granule protein was evaluated for serodiagnosis of Neospora caninum in cattle and water buffalo. NcGRA7, from N. caninum tachyzoites was amplified. PCR product and pMAL-c2X plasmid were digested with EcoR1 restriction enzyme and ex­pressed in Escherichia coli to evaluate its competence for detection of anti- N. cani­num antibodies with ELISA in comparison with commercial IDEXX ELISA. Further­more, 230 sera of presumably healthy cattle and water buffaloes (108 cattle and 122 water buffaloes) were analyzed by both tests to determine the agreement of these two procedures. Sensitivities and specificities of NcGRA7-based ELISA were 94.64% and 90.38% respectively using sera of cattle, but were 98.57% and 86.54% in the case of buffaloes respectively. A good correlation between the results of IDEXX ELISA and ELISA based on recombinant NcGRA7 for detecting N. caninum antibod­ies was appeared. Analyzing by Mc Nemar´s showed that NcGRA7-based ELISA has acceptable capability to differentiate the positive results in comparison with IDEXX ELISA. NcGRA7-based ELISA considering utilized new fragment of ge­nomic DNA is a good tool for serodiagnosis of anti- N. caninum antibodies for screening and epidemiological purposes on cattle herd and water buffaloes as well.

Filamentous hemagglutinin (FHA) is one of the most important immunoprotective antigens of Bordetella pertussis (B.pertussis) and a major component of the acellular pertussis vaccine. In the present study, three overlapping recombinant fragments from the immunodominant region of FHA were produced and their immunogenicity was investigated. Three overlapping coding sequences of FHA antigen were amplified from B.pertussis genomic DNA by PCR. Amplified fragments were expressed in Escherichia coli (E. coli) BL21(DE3) strain and purified through His-tag using Nickel-based chromatography. Purified fragments were characterized by SDS-PAGE and Western blotting techniques. In vitro peripheral blood mononuclear cells (PBMC) proliferation and IFN-γ production were assessed in a limited number of healthy adults vaccinated with a commercial acellular pertussis vaccine in response to all purified FHA fragments by H3-Thymidine incorporation and ELISA, respectively. Recombinant FHA segments were successfully cloned and produced at high levels in E. coli BL21(DE3). SDS-PAGE and Western blot analyses confirmed their purity and reactivity. All three recombinant fragments together with a commercial native FHA were able to induce in vitro PBMC proliferation and IFN-γ production. Our preliminary results suggest that these overlapping recombinant FHA fragments are immunogenic and may prove to be immuno-protective.