In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells
Background: Angiogenesis which occurs mandatory in solid tumors, is a critical step in malignancy progression. Vascular endothelial growth factor (VEGF) is mainly responsible for angiogenesis process and facilitates the formation of new vessels. Distribution of monoclonal antibodies against VEGF or...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2017-01-01
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| Series: | Advanced Biomedical Research |
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| Online Access: | http://www.advbiores.net/article.asp?issn=2277-9175;year=2017;volume=6;issue=1;spage=144;epage=144;aulast=Langari |
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| author | Jahangir Langari Morteza Karimipoor Majid Golkar Hossein Khanahmad Sirous Zeinali Skandar Omidinia Reza Ahangari Cohan Mahdi Behdani Jalal Babaie Roghaye Arezumand Reza Moazami |
| author_facet | Jahangir Langari Morteza Karimipoor Majid Golkar Hossein Khanahmad Sirous Zeinali Skandar Omidinia Reza Ahangari Cohan Mahdi Behdani Jalal Babaie Roghaye Arezumand Reza Moazami |
| author_sort | Jahangir Langari |
| collection | DOAJ |
| description | Background: Angiogenesis which occurs mandatory in solid tumors, is a critical step in malignancy progression. Vascular endothelial growth factor (VEGF) is mainly responsible for angiogenesis process and facilitates the formation of new vessels. Distribution of monoclonal antibodies against VEGF or VEGF receptor (VEGFR) into the solid tumors is limited because of their huge dimensions. Moreover, many investigations have demonstrated the usefulness of immunotoxins to halt angiogenesis in solid tumors. Materials and Methods: We designed, expressed and evaluated the cytotoxicity of a novel nano-immunotoxin composed of VEGF splice variant containing 121 amino acids (VEGF121) and truncated the exotoxin A of Pseudomonas aeruginosa (PE38-KDEL). The fusion protein VEGF121-PE38 was successfully cloned and expressed in Escherichia coli, purified by Ni+ 2 affinity chromatography. The fusion protein was subsequently subjected to refolding using the reduced and oxidized glutathione. Results: The expression level of the fusion protein reached to 1 mg/ml. The VEGF121-PE38 immunotoxin showed a 59 KDa MW which had cytotoxic effect on HUVEC and 293/KDR cells as low and high expressing VEGFR2 cells, respectively. But the cytotoxicity on 293/KDR was 100 folds more than that of VEGFR2 low expressing cell HUVEC. Conclusion: The designed immunotoxin showed more selectivity for higher VEGFR2 expressing cells in vitro. |
| first_indexed | 2024-12-19T18:17:44Z |
| format | Article |
| id | doaj.art-996f91096392477b893248e0e9380d2c |
| institution | Directory Open Access Journal |
| issn | 2277-9175 |
| language | English |
| last_indexed | 2024-12-19T18:17:44Z |
| publishDate | 2017-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Advanced Biomedical Research |
| spelling | doaj.art-996f91096392477b893248e0e9380d2c2022-12-21T20:11:03ZengWolters Kluwer Medknow PublicationsAdvanced Biomedical Research2277-91752017-01-016114414410.4103/2277-9175.218691In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor CellsJahangir LangariMorteza KarimipoorMajid GolkarHossein KhanahmadSirous ZeinaliSkandar OmidiniaReza Ahangari CohanMahdi BehdaniJalal BabaieRoghaye ArezumandReza MoazamiBackground: Angiogenesis which occurs mandatory in solid tumors, is a critical step in malignancy progression. Vascular endothelial growth factor (VEGF) is mainly responsible for angiogenesis process and facilitates the formation of new vessels. Distribution of monoclonal antibodies against VEGF or VEGF receptor (VEGFR) into the solid tumors is limited because of their huge dimensions. Moreover, many investigations have demonstrated the usefulness of immunotoxins to halt angiogenesis in solid tumors. Materials and Methods: We designed, expressed and evaluated the cytotoxicity of a novel nano-immunotoxin composed of VEGF splice variant containing 121 amino acids (VEGF121) and truncated the exotoxin A of Pseudomonas aeruginosa (PE38-KDEL). The fusion protein VEGF121-PE38 was successfully cloned and expressed in Escherichia coli, purified by Ni+ 2 affinity chromatography. The fusion protein was subsequently subjected to refolding using the reduced and oxidized glutathione. Results: The expression level of the fusion protein reached to 1 mg/ml. The VEGF121-PE38 immunotoxin showed a 59 KDa MW which had cytotoxic effect on HUVEC and 293/KDR cells as low and high expressing VEGFR2 cells, respectively. But the cytotoxicity on 293/KDR was 100 folds more than that of VEGFR2 low expressing cell HUVEC. Conclusion: The designed immunotoxin showed more selectivity for higher VEGFR2 expressing cells in vitro.http://www.advbiores.net/article.asp?issn=2277-9175;year=2017;volume=6;issue=1;spage=144;epage=144;aulast=LangariImmunotoxinpseudomonas exotoxin Asolid tumorvascular endothelial growth factor |
| spellingShingle | Jahangir Langari Morteza Karimipoor Majid Golkar Hossein Khanahmad Sirous Zeinali Skandar Omidinia Reza Ahangari Cohan Mahdi Behdani Jalal Babaie Roghaye Arezumand Reza Moazami In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells Advanced Biomedical Research Immunotoxin pseudomonas exotoxin A solid tumor vascular endothelial growth factor |
| title | In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells |
| title_full | In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells |
| title_fullStr | In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells |
| title_full_unstemmed | In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells |
| title_short | In Vitro Evaluation of Vegf-Pseudomonas Exotoxin: A Conjugated on Tumor Cells |
| title_sort | in vitro evaluation of vegf pseudomonas exotoxin a conjugated on tumor cells |
| topic | Immunotoxin pseudomonas exotoxin A solid tumor vascular endothelial growth factor |
| url | http://www.advbiores.net/article.asp?issn=2277-9175;year=2017;volume=6;issue=1;spage=144;epage=144;aulast=Langari |
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