schliessen

Filtern

 

Bibliotheken

Production and characterization of a CD25-specific scFv-Fc antibody secreted from Pichia pastoris

Antibodies against CD25 would be novel tools for the diagnosis and treatment of adult T cell leukemia lymphoma (ATLL) and many other immune disorders. In our previous work, we successfully produced the single-chain fragment of a variable antibody against CD25, the Dmab(scFv) antibody, using Pichia p... Full description

Journal Title: Applied Microbiology and Biotechnology 2013, Vol.97(9), pp.3855-3863
Main Author: Wan, Lin
Other Authors: Zhu, Shengyun , Zhu, Jian , Yang, Hao , Li, Shengfu , Li, Youping , Cheng, Jingqiu , Lu, Xiaofeng
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 0175-7598 ; E-ISSN: 1432-0614 ; DOI: 10.1007/s00253-012-4632-9
Link: http://dx.doi.org/10.1007/s00253-012-4632-9
Zum Text:
SendSend as email Add to Book BagAdd to Book Bag
Staff View
recordid: springer_jour10.1007/s00253-012-4632-9
title: Production and characterization of a CD25-specific scFv-Fc antibody secreted from Pichia pastoris
format: Article
creator:
  • Wan, Lin
  • Zhu, Shengyun
  • Zhu, Jian
  • Yang, Hao
  • Li, Shengfu
  • Li, Youping
  • Cheng, Jingqiu
  • Lu, Xiaofeng
subjects:
  • Immunotherapy
  • Single-chain variable fragment
  • Antibody engineering
  • CD25
  • P. pastoris
ispartof: Applied Microbiology and Biotechnology, 2013, Vol.97(9), pp.3855-3863
description: Antibodies against CD25 would be novel tools for the diagnosis and treatment of adult T cell leukemia lymphoma (ATLL) and many other immune disorders. In our previous work, we successfully produced the single-chain fragment of a variable antibody against CD25, the Dmab(scFv) antibody, using Pichia pastoris . Here, we describe a novel form of an antibody against CD25, the Dmab(scFv)-Fc antibody, also produced by P. pastoris . To construct the Dmab(scFv)-Fc antibody, the Dmab(scFv) antibody was genetically fused to the Fc fragment of a human IgG1 antibody. A fusion gene encoding Dmab(scFv)-Fc antibody was cloned into the pPIC9K plasmid and expressed at high levels, 60–70 mg/l, by P. pastoris under optimized conditions. The Dmab(scFv)-Fc antibody was similar to the Dmab(scFv) antibody in its binding specificity but different in its molecular form and Fc-mediated effector functions. The Dmab(scFv)-Fc antibody and the Dmab(scFv) antibody both bound to CD25-positive MJ cells but not to CD25-negative K562 cells. The Dmab(scFv)-Fc antibody existed as a dimer whereas the Dmab(scFv) antibody was a monomer because it lacks the Fc fragment. The Dmab(scFv)-Fc antibody enhanced the antibody-dependent cellular cytotoxicity of CD25-positive cancer cells, whereas the Dmab(scFv) antibody was inactive in the antibody-dependent cellular cytotoxicity assays. In addition, compared to the Dmab(scFv) antibody, the Dmab(scFv)-Fc antibody showed stronger immunosuppressive activity in the Con A-stimulated lymphocyte proliferation system and in the mixed lymphocyte reaction system. These results demonstrate that the Dmab(scFv)-Fc antibody produced in P. pastoris is functional, and therefore it might be developed as a novel diagnostic and therapeutic tool for ATLL and other immune disorders.
language: eng
source:
identifier: ISSN: 0175-7598 ; E-ISSN: 1432-0614 ; DOI: 10.1007/s00253-012-4632-9
fulltext: fulltext
issn:
  • 1432-0614
  • 14320614
  • 0175-7598
  • 01757598
url: Link


@attributes
ID927645445
RANK0.06999999
NO1
SEARCH_ENGINEprimo_central_multiple_fe
SEARCH_ENGINE_TYPEPrimo Central Search Engine
LOCALfalse
PrimoNMBib
record
control
sourcerecordid10.1007/s00253-012-4632-9
sourceidspringer_jour
recordidTN_springer_jour10.1007/s00253-012-4632-9
sourcesystemPC
pqid1352286143
galeid337618222
display
typearticle
titleProduction and characterization of a CD25-specific scFv-Fc antibody secreted from Pichia pastoris
creatorWan, Lin ; Zhu, Shengyun ; Zhu, Jian ; Yang, Hao ; Li, Shengfu ; Li, Youping ; Cheng, Jingqiu ; Lu, Xiaofeng
ispartofApplied Microbiology and Biotechnology, 2013, Vol.97(9), pp.3855-3863
identifier
subjectImmunotherapy ; Single-chain variable fragment ; Antibody engineering ; CD25 ; P. pastoris
descriptionAntibodies against CD25 would be novel tools for the diagnosis and treatment of adult T cell leukemia lymphoma (ATLL) and many other immune disorders. In our previous work, we successfully produced the single-chain fragment of a variable antibody against CD25, the Dmab(scFv) antibody, using Pichia pastoris . Here, we describe a novel form of an antibody against CD25, the Dmab(scFv)-Fc antibody, also produced by P. pastoris . To construct the Dmab(scFv)-Fc antibody, the Dmab(scFv) antibody was genetically fused to the Fc fragment of a human IgG1 antibody. A fusion gene encoding Dmab(scFv)-Fc antibody was cloned into the pPIC9K plasmid and expressed at high levels, 60–70 mg/l, by P. pastoris under optimized conditions. The Dmab(scFv)-Fc antibody was similar to the Dmab(scFv) antibody in its binding specificity but different in its molecular form and Fc-mediated effector functions. The Dmab(scFv)-Fc antibody and the Dmab(scFv) antibody both bound to CD25-positive MJ cells but not to CD25-negative K562 cells. The Dmab(scFv)-Fc antibody existed as a dimer whereas the Dmab(scFv) antibody was a monomer because it lacks the Fc fragment. The Dmab(scFv)-Fc antibody enhanced the antibody-dependent cellular cytotoxicity of CD25-positive cancer cells, whereas the Dmab(scFv) antibody was inactive in the antibody-dependent cellular cytotoxicity assays. In addition, compared to the Dmab(scFv) antibody, the Dmab(scFv)-Fc antibody showed stronger immunosuppressive activity in the Con A-stimulated lymphocyte proliferation system and in the mixed lymphocyte reaction system. These results demonstrate that the Dmab(scFv)-Fc antibody produced in P. pastoris is functional, and therefore it might be developed as a novel diagnostic and therapeutic tool for ATLL and other immune disorders.
languageeng
source
version8
lds50peer_reviewed
links
openurl$$Topenurl_article
openurlfulltext$$Topenurlfull_article
backlink$$Uhttp://dx.doi.org/10.1007/s00253-012-4632-9$$EView_full_text_in_Springer_(Subscribers_only)
search
creatorcontrib
0Wan, Lin
1Zhu, Shengyun
2Zhu, Jian
3Yang, Hao
4Li, Shengfu
5Li, Youping
6Cheng, Jingqiu
7Lu, Xiaofeng
titleProduction and characterization of a CD25-specific scFv-Fc antibody secreted from Pichia pastoris
descriptionAntibodies against CD25 would be novel tools for the diagnosis and treatment of adult T cell leukemia lymphoma (ATLL) and many other immune disorders. In our previous work, we successfully produced the single-chain fragment of a variable antibody against CD25, the Dmab(scFv) antibody, using Pichia pastoris . Here, we describe a novel form of an antibody against CD25, the Dmab(scFv)-Fc antibody, also produced by P. pastoris . To construct the Dmab(scFv)-Fc antibody, the Dmab(scFv) antibody was genetically fused to the Fc fragment of a human IgG1 antibody. A fusion gene encoding Dmab(scFv)-Fc antibody was cloned into the pPIC9K plasmid and expressed at high levels, 60–70 mg/l, by P. pastoris under optimized conditions. The Dmab(scFv)-Fc antibody was similar to the Dmab(scFv) antibody in its binding specificity but different in its molecular form and Fc-mediated effector functions. The Dmab(scFv)-Fc antibody and the Dmab(scFv) antibody both bound to CD25-positive MJ cells but not to CD25-negative K562 cells. The Dmab(scFv)-Fc antibody existed as a dimer whereas the Dmab(scFv) antibody was a monomer because it lacks the Fc fragment. The Dmab(scFv)-Fc antibody enhanced the antibody-dependent cellular cytotoxicity of CD25-positive cancer cells, whereas the Dmab(scFv) antibody was inactive in the antibody-dependent cellular cytotoxicity assays. In addition, compared to the Dmab(scFv) antibody, the Dmab(scFv)-Fc antibody showed stronger immunosuppressive activity in the Con A-stimulated lymphocyte proliferation system and in the mixed lymphocyte reaction system. These results demonstrate that the Dmab(scFv)-Fc antibody produced in P. pastoris is functional, and therefore it might be developed as a novel diagnostic and therapeutic tool for ATLL and other immune disorders.
subject
0Immunotherapy
1Single-chain variable fragment
2Antibody engineering
3CD25
4P. pastoris
general
010.1007/s00253-012-4632-9
1English
2Springer Science & Business Media B.V.
3SpringerLink
sourceidspringer_jour
recordidspringer_jour10.1007/s00253-012-4632-9
issn
01432-0614
114320614
20175-7598
301757598
rsrctypearticle
creationdate2013
addtitle
0Applied Microbiology and Biotechnology
1Appl Microbiol Biotechnol
searchscopespringer_journals_complete
scopespringer_journals_complete
lsr30VSR-Enriched:[pqid, galeid, pages]
sort
titleProduction and characterization of a CD25-specific scFv-Fc antibody secreted from Pichia pastoris
authorWan, Lin ; Zhu, Shengyun ; Zhu, Jian ; Yang, Hao ; Li, Shengfu ; Li, Youping ; Cheng, Jingqiu ; Lu, Xiaofeng
creationdate20130500
facets
frbrgroupid9138880650641431239
frbrtype5
languageeng
creationdate2013
topic
0Immunotherapy
1Single-Chain Variable Fragment
2Antibody Engineering
3Cd25
4P. Pastoris
collectionSpringerLink
prefilterarticles
rsrctypearticles
creatorcontrib
0Wan, Lin
1Zhu, Shengyun
2Zhu, Jian
3Yang, Hao
4Li, Shengfu
5Li, Youping
6Cheng, Jingqiu
7Lu, Xiaofeng
jtitleApplied Microbiology And Biotechnology
toplevelpeer_reviewed
delivery
delcategoryRemote Search Resource
fulltextfulltext
addata
aulast
0Wan
1Zhu
2Yang
3Li
4Cheng
5Lu
aufirst
0Lin
1Shengyun
2Jian
3Hao
4Shengfu
5Youping
6Jingqiu
7Xiaofeng
au
0Wan, Lin
1Zhu, Shengyun
2Zhu, Jian
3Yang, Hao
4Li, Shengfu
5Li, Youping
6Cheng, Jingqiu
7Lu, Xiaofeng
atitleProduction and characterization of a CD25-specific scFv-Fc antibody secreted from Pichia pastoris
jtitleApplied Microbiology and Biotechnology
stitleAppl Microbiol Biotechnol
risdate201305
volume97
issue9
spage3855
epage3863
issn0175-7598
eissn1432-0614
genrearticle
ristypeJOUR
abstractAntibodies against CD25 would be novel tools for the diagnosis and treatment of adult T cell leukemia lymphoma (ATLL) and many other immune disorders. In our previous work, we successfully produced the single-chain fragment of a variable antibody against CD25, the Dmab(scFv) antibody, using Pichia pastoris . Here, we describe a novel form of an antibody against CD25, the Dmab(scFv)-Fc antibody, also produced by P. pastoris . To construct the Dmab(scFv)-Fc antibody, the Dmab(scFv) antibody was genetically fused to the Fc fragment of a human IgG1 antibody. A fusion gene encoding Dmab(scFv)-Fc antibody was cloned into the pPIC9K plasmid and expressed at high levels, 60–70 mg/l, by P. pastoris under optimized conditions. The Dmab(scFv)-Fc antibody was similar to the Dmab(scFv) antibody in its binding specificity but different in its molecular form and Fc-mediated effector functions. The Dmab(scFv)-Fc antibody and the Dmab(scFv) antibody both bound to CD25-positive MJ cells but not to CD25-negative K562 cells. The Dmab(scFv)-Fc antibody existed as a dimer whereas the Dmab(scFv) antibody was a monomer because it lacks the Fc fragment. The Dmab(scFv)-Fc antibody enhanced the antibody-dependent cellular cytotoxicity of CD25-positive cancer cells, whereas the Dmab(scFv) antibody was inactive in the antibody-dependent cellular cytotoxicity assays. In addition, compared to the Dmab(scFv) antibody, the Dmab(scFv)-Fc antibody showed stronger immunosuppressive activity in the Con A-stimulated lymphocyte proliferation system and in the mixed lymphocyte reaction system. These results demonstrate that the Dmab(scFv)-Fc antibody produced in P. pastoris is functional, and therefore it might be developed as a novel diagnostic and therapeutic tool for ATLL and other immune disorders.
copBerlin/Heidelberg
pubSpringer-Verlag
doi10.1007/s00253-012-4632-9
pages3855-3863
date2013-05