Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination
Aim: Idiotype, the unique part of immunoglobulin molecule expressed on the surface of B-cells, represents a specific antigen for vaccination against lymphoma. We have developed a rapid method for immunoglobulin variable fragments cloning, assembling and expression of recombinant idiotype protein in...
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| Опубліковано в: : | Experimental Oncology |
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| Дата: | 2013 |
| Автори: | , , , , , |
| Формат: | Стаття |
| Мова: | Англійська |
| Опубліковано: |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2013
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| Онлайн доступ: | https://nasplib.isofts.kiev.ua/handle/123456789/139103 |
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| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination / A.N. Meleshko, K.P. Vashkevich, E.G. Fomina, E.P. Scheslenok, T.V. Schkolina, G.V. Sergeev // Experimental Oncology. — 2013. — Т. 35, № 1. — С. 8-14. — Бібліогр.: 34 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1862628316021784576 |
|---|---|
| author | Meleshko, A.N. Vashkevich, K.P. Fomina, E.G. Scheslenok, E.P. Schkolina, T.V. Sergeev, G.V. |
| author_facet | Meleshko, A.N. Vashkevich, K.P. Fomina, E.G. Scheslenok, E.P. Schkolina, T.V. Sergeev, G.V. |
| citation_txt | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination / A.N. Meleshko, K.P. Vashkevich, E.G. Fomina, E.P. Scheslenok, T.V. Schkolina, G.V. Sergeev // Experimental Oncology. — 2013. — Т. 35, № 1. — С. 8-14. — Бібліогр.: 34 назв. — англ. |
| collection | DSpace DC |
| container_title | Experimental Oncology |
| description | Aim: Idiotype, the unique part of immunoglobulin molecule expressed on the surface of B-cells, represents a specific antigen for vaccination against lymphoma. We have developed a rapid method for immunoglobulin variable fragments cloning, assembling and expression of recombinant idiotype protein in Escherichia coli. Methods: PCR with specially designed panel of primers was used for direct amplification of variable regions of tumor immunoglobulin. Overlapping extension PCR, restriction and ligation was applied for assembling and cloning of vaccine construction. Idiotype protein was purified by metal-chelate chromatography. Results: Methods of idiotype cloning from lymphoma cells and production of recombinant protein were developed and optimized. Several samples of idiotypic proteins originating from B-cell lines and lymphoma patients were produced. Conclusion: The proposed method of vaccine production is relatively cheap, not very laborious and requires as long as 6–7 week to perform. The expressed protein was soluble, did not accumulate in inclusion bodies and harvested at sufficient for vaccination quantity and concentration.
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| first_indexed | 2025-11-30T09:12:33Z |
| format | Article |
| fulltext | |
| id | nasplib_isofts_kiev_ua-123456789-139103 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-11-30T09:12:33Z |
| publishDate | 2013 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Meleshko, A.N. Vashkevich, K.P. Fomina, E.G. Scheslenok, E.P. Schkolina, T.V. Sergeev, G.V. 2018-06-19T19:09:55Z 2018-06-19T19:09:55Z 2013 Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination / A.N. Meleshko, K.P. Vashkevich, E.G. Fomina, E.P. Scheslenok, T.V. Schkolina, G.V. Sergeev // Experimental Oncology. — 2013. — Т. 35, № 1. — С. 8-14. — Бібліогр.: 34 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/139103 Aim: Idiotype, the unique part of immunoglobulin molecule expressed on the surface of B-cells, represents a specific antigen for vaccination against lymphoma. We have developed a rapid method for immunoglobulin variable fragments cloning, assembling and expression of recombinant idiotype protein in Escherichia coli. Methods: PCR with specially designed panel of primers was used for direct amplification of variable regions of tumor immunoglobulin. Overlapping extension PCR, restriction and ligation was applied for assembling and cloning of vaccine construction. Idiotype protein was purified by metal-chelate chromatography. Results: Methods of idiotype cloning from lymphoma cells and production of recombinant protein were developed and optimized. Several samples of idiotypic proteins originating from B-cell lines and lymphoma patients were produced. Conclusion: The proposed method of vaccine production is relatively cheap, not very laborious and requires as long as 6–7 week to perform. The expressed protein was soluble, did not accumulate in inclusion bodies and harvested at sufficient for vaccination quantity and concentration. This work was supported by the Health Ministry
 of Belarus (Grant # 20101848) and The Foundation
 for Fundamental Research, National Academy of Sciences
 of Belarus (Grunt # 20081881). en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination Article published earlier |
| spellingShingle | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination Meleshko, A.N. Vashkevich, K.P. Fomina, E.G. Scheslenok, E.P. Schkolina, T.V. Sergeev, G.V. Original contributions |
| title | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination |
| title_full | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination |
| title_fullStr | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination |
| title_full_unstemmed | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination |
| title_short | Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination |
| title_sort | cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human scfv protein in e. coli for anti-idiotype vaccination |
| topic | Original contributions |
| topic_facet | Original contributions |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/139103 |
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