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Seminario Biomol 2023.pptx
- 1. Estefanía Zapata Rúa Tercer semestre Universidad pontificia bolivariana
- 2. INTRODUCTION Hepatocellular carcinoma (HCC) is the seventh most prevalent liver cancer in the world and the fourth leading cause of cancer mortality worldwide. The carcinoma consists of epithelial cells, which are the cells that line the inner and outer parts of the body. Treatments against this type of cancer are scarce and it depends on the stage of the cancer. Patients can be treated with surgery or chemotherapy with sorafenib.
- 3. INTRODUCTION Researches have used adoptive immunotherapy as indirect immunological approaches including vaccines. Vaccines are an effective strategy against tumor cells and are generally divided into three types, including pulsed dendritic cells (DCs), peptide-based vaccines, and DNA-based vaccines. Multi-epitope peptide vaccines are used against pathogenic bacteria, viruses, and tumor cells.
- 4. General Objetive Production of a Novel Multi-Epitope Peptide Vaccine against Hepatocellular Carcinoma
- 5. Designed Vaccine Construct Expression of the Vaccine Protein Materials and Methods It was used bioinformatical servers (IEDB, ProPred-I PAComplex and PAComplex ) for the design of a novel multi-epitope peptide vaccine against HCC. The media culture of E. coli was at 37° and 1 mL was diluted in ampicillin. Then it was shaken induced via two different concentrations of (IPTG). This process was to transform the cells via centrifugation and the obtained pellets
- 6. Western Blot Test SDS-PAGE Análisis Materials and Methods It was used to prove the expresión of the vaccine. This is a technique used to identify a specific protein. The method involves the use of gel electrophoresis to separate the proteins in the sample. It is used to separate proteins according to their electrophoretic mobility. It is widely used to quantify, compare and characterize proteins. Constitutes the input technique for the performance of Western Blotting
- 7. Results The SDS-PAGE technique allowed to analyses of the multi-epitope peptide expression of the vaccine. The result of SDS-PAGE analysis of the multi-epitope peptide vaccine purification.
- 8. Results The Western blot analysis results.
- 9. Author What the autor says Agree or disagree Sawada Y, Yoshikawa T, Ofuji K, Yoshimura M, Tsuchiya N, Takahashi M The linear and conformational B-cell epitopes and interferon-gamma-inducing epitopes were recognized in the vaccine construct, because provoking the humoral and cellular immune responses is essential in multiepitope peptide vaccines.3 Agree Li J, Xing Y, Zhou Z, Yao W, Cao R, Li T HCC is a cancer with a high mortality rate. This malignancy is not usually diagnosed in the primary stages, because it is known as a heterogeneous disorder that includes several etiological agents; therefore, it is diagnosed in the advanced stages, when it is usually late for treatment. Agree Gravel P, Golaz O. To prove the expression and purification oft he recombinant vaccine protein, the Western blot method was employed. Agree Discussions
- 10. Conclusions E. coli is an acceptable organism to represent recombinant proteins. This prokaryotic cell allowed acceptable implementation techniques for producing multi-epitope peptide vaccine against HCC Computer tools were of great help in creating a novel multi-epitope peptide vaccine against HCC
Editor's Notes
- Treatments against this type of cancer are scarce and it depends on the stage of the cancer. Patients can be treated with surgery (Primary Phase) or chemotherapy with sorafenib. But only 10% of patients survive more than three years.
- In recent years, researchers have used three main approaches to stimulate the immune response against HCC, including adoptive immunotherapy as indirect immunological approaches including vaccines. Vaccines, which belong to the indirect immunological strategies, are an effective strategy against tumor cells and are generally divided into three types, including pulsed dendritic cells (DCs), peptide-based vaccines, and DNA-based vaccines. Research has shown Multi-epitope peptide vaccines are used against pathogenic bacteria, viruses, and tumor cells. This consist of Cytotoxic T cell Lymphocytes (CTL) and Helper T cell Lymphocytes (HTL) epitopes, and provoke humoral and adaptive immune responses, these are considered as a beneficial treatment for tumor immunotherapy
- It was used bioinformatical servers (IEDB, ProPred-I PAComplex and PAComplex ) for the design of a novel multi-epitope peptide vaccine against HCC. It includes building peptides as MHC class-I peptides. The media culture of E. coli was at 37° overnight. After that, 1 mL was diluted in ampicillin. Then it was shaken until an optical Density. The culture was induced via two different concentrations of Isopropyl β-D-1-thiogalactopyranoside (IPTG). The transformed cells were harvested via centrifugation and the obtained pellets were stored at -20.
- It is used to separate proteins according to their electrophoretic mobility. It is widely used to quantify, compare and characterize proteins. Constitutes the input technique for the performance of Western Blotting It was used to prove the expresión of the vaccine. This is a technique used to identify a specific protein. The method involves the use of gel electrophoresis to separate the proteins in the sample. Then the proteins are transferred to a membrane which exposes an antibody against the protein. Functional binding is detected using a radioactive or chemical tracer.
- The SDS-PAGE technique allowed to analyses of the multi-epitope peptide expression of the vaccine. The line 2 shows the expressed protein before expression induction, and lines 3-6 show the protein expression after Expression. The black circle represents the optimal protein expression. The result of SDS-PAGE analysis of the multi-epitope peptide vaccine purification. Line 1 shows transformed E. coli after expression. lines 2-3 show the unpurified peptide, and line 4 shows the purified peptide vaccine.
- Line 1 shows the purified protein using the Western blot technique.