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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Published in:Experimental Oncology
Date:2013
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
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Original contributions
Online Access:https://nasplib.isofts.kiev.ua/handle/123456789/139103
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Journal Title:Digital Library of Periodicals of National Academy of Sciences of Ukraine
Cite this: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.

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Digital Library of Periodicals of National Academy of Sciences of Ukraine
id nasplib_isofts_kiev_ua-123456789-139103
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
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
title Cloning of variable fragments of tumor immunoglobulin, assembling and expressing of human SCFV protein in E. coli for anti-idiotype vaccination
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_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
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.
topic Original contributions
topic_facet Original contributions
publishDate 2013
language English
container_title Experimental Oncology
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
format Article
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.
issn 1812-9269
url https://nasplib.isofts.kiev.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.
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first_indexed 2025-11-30T09:12:33Z
last_indexed 2025-11-30T09:12:33Z
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