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Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination

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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Main Authors: Meleshko, A.N., Vashkevich, K.P., Fomina, E.G., Scheslenok, E.P., Schkolina, T.V., Sergeev, G.V.
Format: Article
Language:English
Published: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2013
Series:Experimental Oncology
Subjects:
Original contributions
Online Access:http://dspace.nbuv.gov.ua/handle/123456789/139103
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spelling irk-123456789-1391032018-06-20T03:10:15Z 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 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. 2013 Article 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 http://dspace.nbuv.gov.ua/handle/123456789/139103 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
language English
topic Original contributions
Original contributions
spellingShingle Original contributions
Original contributions
Meleshko, A.N.
Vashkevich, K.P.
Fomina, E.G.
Scheslenok, E.P.
Schkolina, T.V.
Sergeev, G.V.
Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination
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.
format Article
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.
author_sort Meleshko, A.N.
title 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_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_sort cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human scfv protein in e. coli for anti-idiotype vaccination
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
publishDate 2013
topic_facet Original contributions
url http://dspace.nbuv.gov.ua/handle/123456789/139103
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.
series Experimental Oncology
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first_indexed 2023-10-18T21:19:37Z
last_indexed 2023-10-18T21:19:37Z
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