Conversion of murine Fabs isolated from a combinatorial phage display library to full length immunoglobulins
Identifieur interne : 002B11 ( Main/Exploration ); précédent : 002B10; suivant : 002B12Conversion of murine Fabs isolated from a combinatorial phage display library to full length immunoglobulins
Auteurs : Robert S. Ames [États-Unis] ; Mark A. Tornetta [États-Unis] ; Keith Deen [États-Unis] ; Christopher S. Jones [États-Unis] ; Ann M. Swift [États-Unis] ; Subinay Ganguly [États-Unis]Source :
- Journal of Immunological Methods [ 0022-1759 ] ; 1995.
English descriptors
- Teeft :
- Acad, Ames, Assay buffer, Bacterial neomycin phosphotransferase gene, Binding activity, Cdna, Cdnas coding, Cell lines, Cell membranes, Cell supernatants, Cell surface antigen, Chain disulfide bond, Coding region, Combinatorial, Combinatorial libraries, Combinatorial library, Combinatorial library construction, Constant domain, Distress syndrome, Elisa, Entire coding sequence, Expression cassette, Expression vectors, Fabs, Fabs reactive, Fragment, Full length, Full length igg2a, Genetic engineering, Gradient polyacrylamide, Heavy chain, Heavy chain expression vector, Human fabs, Human immunodeficiency virus, Igg2a, Igg2a mabs, Igg2a myeloma protein, Immunological, Immunological methods, Light chain, Light chain expression vector, Light chains, Mabs, Malarial infection, Mammalian cells, Mammalian expression vectors, Microtiter wells, Monoclonal, Monoclonal antibodies, Monoclonal antibody, Murine, Murine fabs, Natl, Part ligation, Phage, Plasmid, Plasmid muhcosint, Plasmid mulcosint, Polik, Proc, Quickmab system, Rc5a, Respiratory syncytial virus, Resultant plasmid, Room temperature, Single step, Single step conversion, Smithkline beecham pharmaceuticals, Stable cell lines, Stuffer fragment, Xhoi site.
Abstract
Abstract: The use of combinatorial Ig libraries displayed on the surface of bacteriophage has advantages over traditional hybridoma techniques for the generation of mAbs but in many instances full length Igs may be more desirable than Fab fragments. Two murine Fabs reactive with the human complement component C5a, recovered from a combinatorial library, were converted to full length IgG2a mAbs. The VH and VL domains of these antibodies were removed from the bacterial expression vector used for the combinatorial library construction, and subcloned into individual mammalian expression vectors containing the corresponding Ig heavy and light chain constant regions. The subcloning relied on 5′ restriction endonuclease sites encoded by the oligonucleotide primers originally used to amplify the Ig cDNAs and 3′ sites conserved in CH1 and Cκ. These vectors were co-transfected into COS cells yielding full length IgG2a versions of the anti-C5a antibodies. The mAbs, purified from the culture supernatant, retained the full activity of the Fabs, binding specifically to and neutralizing human recombinant C5a. Refined versions of the mammalian expression vectors have been constructed for single step conversion of murine recombinant Fabs, recovered from combinatorial libraries, to IgG2a mAbs.
Url:
DOI: 10.1016/0022-1759(95)00086-P
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001B98
- to stream Istex, to step Curation: 001B98
- to stream Istex, to step Checkpoint: 001899
- to stream Main, to step Merge: 002B67
- to stream Main, to step Curation: 002B11
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Conversion of murine Fabs isolated from a combinatorial phage display library to full length immunoglobulins</title><author><name sortKey="Ames, Robert S" sort="Ames, Robert S" uniqKey="Ames R" first="Robert S." last="Ames">Robert S. Ames</name></author><author><name sortKey="Tornetta, Mark A" sort="Tornetta, Mark A" uniqKey="Tornetta M" first="Mark A." last="Tornetta">Mark A. Tornetta</name></author><author><name sortKey="Deen, Keith" sort="Deen, Keith" uniqKey="Deen K" first="Keith" last="Deen">Keith Deen</name></author><author><name sortKey="Jones, Christopher S" sort="Jones, Christopher S" uniqKey="Jones C" first="Christopher S." last="Jones">Christopher S. Jones</name></author><author><name sortKey="Swift, Ann M" sort="Swift, Ann M" uniqKey="Swift A" first="Ann M." last="Swift">Ann M. Swift</name></author><author><name sortKey="Ganguly, Subinay" sort="Ganguly, Subinay" uniqKey="Ganguly S" first="Subinay" last="Ganguly">Subinay Ganguly</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:70235285E69BC29D17E92D92C01225B0700E4E8C</idno><date when="1995" year="1995">1995</date><idno type="doi">10.1016/0022-1759(95)00086-P</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-5VCB9B2V-N/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001B98</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001B98</idno><idno type="wicri:Area/Istex/Curation">001B98</idno><idno type="wicri:Area/Istex/Checkpoint">001899</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001899</idno><idno type="wicri:doubleKey">0022-1759:1995:Ames R:conversion:of:murine</idno><idno type="wicri:Area/Main/Merge">002B67</idno><idno type="wicri:Area/Main/Curation">002B11</idno><idno type="wicri:Area/Main/Exploration">002B11</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Conversion of murine Fabs isolated from a combinatorial phage display library to full length immunoglobulins</title><author><name sortKey="Ames, Robert S" sort="Ames, Robert S" uniqKey="Ames R" first="Robert S." last="Ames">Robert S. Ames</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Molecular Immunology, UE0548, SmithKline Beecham Pharmaceuticals, P.O. Box 1539, King of Prussia, PA 19406-0939</wicri:regionArea><placeName><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Tornetta, Mark A" sort="Tornetta, Mark A" uniqKey="Tornetta M" first="Mark A." last="Tornetta">Mark A. Tornetta</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Molecular Immunology, UE0548, SmithKline Beecham Pharmaceuticals, P.O. Box 1539, King of Prussia, PA 19406-0939</wicri:regionArea><placeName><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Deen, Keith" sort="Deen, Keith" uniqKey="Deen K" first="Keith" last="Deen">Keith Deen</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Molecular Immunology, UE0548, SmithKline Beecham Pharmaceuticals, P.O. Box 1539, King of Prussia, PA 19406-0939</wicri:regionArea><placeName><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Jones, Christopher S" sort="Jones, Christopher S" uniqKey="Jones C" first="Christopher S." last="Jones">Christopher S. Jones</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Protein Biochemistry, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406-0939</wicri:regionArea><placeName><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Swift, Ann M" sort="Swift, Ann M" uniqKey="Swift A" first="Ann M." last="Swift">Ann M. Swift</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Gene Expression Sciences, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406-0939</wicri:regionArea><placeName><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Ganguly, Subinay" sort="Ganguly, Subinay" uniqKey="Ganguly S" first="Subinay" last="Ganguly">Subinay Ganguly</name><affiliation wicri:level="2"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Gene Expression Sciences, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406-0939</wicri:regionArea><placeName><region type="state">Pennsylvanie</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Immunological Methods</title><title level="j" type="abbrev">JIM</title><idno type="ISSN">0022-1759</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1995">1995</date><biblScope unit="volume">184</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="177">177</biblScope><biblScope unit="page" to="186">186</biblScope></imprint><idno type="ISSN">0022-1759</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0022-1759</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acad</term><term>Ames</term><term>Assay buffer</term><term>Bacterial neomycin phosphotransferase gene</term><term>Binding activity</term><term>Cdna</term><term>Cdnas coding</term><term>Cell lines</term><term>Cell membranes</term><term>Cell supernatants</term><term>Cell surface antigen</term><term>Chain disulfide bond</term><term>Coding region</term><term>Combinatorial</term><term>Combinatorial libraries</term><term>Combinatorial library</term><term>Combinatorial library construction</term><term>Constant domain</term><term>Distress syndrome</term><term>Elisa</term><term>Entire coding sequence</term><term>Expression cassette</term><term>Expression vectors</term><term>Fabs</term><term>Fabs reactive</term><term>Fragment</term><term>Full length</term><term>Full length igg2a</term><term>Genetic engineering</term><term>Gradient polyacrylamide</term><term>Heavy chain</term><term>Heavy chain expression vector</term><term>Human fabs</term><term>Human immunodeficiency virus</term><term>Igg2a</term><term>Igg2a mabs</term><term>Igg2a myeloma protein</term><term>Immunological</term><term>Immunological methods</term><term>Light chain</term><term>Light chain expression vector</term><term>Light chains</term><term>Mabs</term><term>Malarial infection</term><term>Mammalian cells</term><term>Mammalian expression vectors</term><term>Microtiter wells</term><term>Monoclonal</term><term>Monoclonal antibodies</term><term>Monoclonal antibody</term><term>Murine</term><term>Murine fabs</term><term>Natl</term><term>Part ligation</term><term>Phage</term><term>Plasmid</term><term>Plasmid muhcosint</term><term>Plasmid mulcosint</term><term>Polik</term><term>Proc</term><term>Quickmab system</term><term>Rc5a</term><term>Respiratory syncytial virus</term><term>Resultant plasmid</term><term>Room temperature</term><term>Single step</term><term>Single step conversion</term><term>Smithkline beecham pharmaceuticals</term><term>Stable cell lines</term><term>Stuffer fragment</term><term>Xhoi site</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The use of combinatorial Ig libraries displayed on the surface of bacteriophage has advantages over traditional hybridoma techniques for the generation of mAbs but in many instances full length Igs may be more desirable than Fab fragments. Two murine Fabs reactive with the human complement component C5a, recovered from a combinatorial library, were converted to full length IgG2a mAbs. The VH and VL domains of these antibodies were removed from the bacterial expression vector used for the combinatorial library construction, and subcloned into individual mammalian expression vectors containing the corresponding Ig heavy and light chain constant regions. The subcloning relied on 5′ restriction endonuclease sites encoded by the oligonucleotide primers originally used to amplify the Ig cDNAs and 3′ sites conserved in CH1 and Cκ. These vectors were co-transfected into COS cells yielding full length IgG2a versions of the anti-C5a antibodies. The mAbs, purified from the culture supernatant, retained the full activity of the Fabs, binding specifically to and neutralizing human recombinant C5a. Refined versions of the mammalian expression vectors have been constructed for single step conversion of murine recombinant Fabs, recovered from combinatorial libraries, to IgG2a mAbs.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Pennsylvanie</li></region></list><tree><country name="États-Unis"><region name="Pennsylvanie"><name sortKey="Ames, Robert S" sort="Ames, Robert S" uniqKey="Ames R" first="Robert S." last="Ames">Robert S. Ames</name></region><name sortKey="Deen, Keith" sort="Deen, Keith" uniqKey="Deen K" first="Keith" last="Deen">Keith Deen</name><name sortKey="Ganguly, Subinay" sort="Ganguly, Subinay" uniqKey="Ganguly S" first="Subinay" last="Ganguly">Subinay Ganguly</name><name sortKey="Jones, Christopher S" sort="Jones, Christopher S" uniqKey="Jones C" first="Christopher S." last="Jones">Christopher S. Jones</name><name sortKey="Swift, Ann M" sort="Swift, Ann M" uniqKey="Swift A" first="Ann M." last="Swift">Ann M. Swift</name><name sortKey="Tornetta, Mark A" sort="Tornetta, Mark A" uniqKey="Tornetta M" first="Mark A." last="Tornetta">Mark A. Tornetta</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/ChloroquineV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002B11 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 002B11 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= ChloroquineV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:70235285E69BC29D17E92D92C01225B0700E4E8C
|texte= Conversion of murine Fabs isolated from a combinatorial phage display library to full length immunoglobulins
}}
| This area was generated with Dilib version V0.6.33. |  |