Identification of phage antibodies toward the Werner protein by selection on Western blots

Peter Ravn; Svend Kjaer; Kristian Hobolt Jensen; T Wind; K B Jensen; P Kristensen; R M Brosh; D K Orren; V A Bohr; B F Clark

doi:10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.0.CO;2-5

Identification of phage antibodies toward the Werner protein by selection on Western blots

Peter Ravn, Svend Kjaer, Kristian Hobolt Jensen, T Wind, K B Jensen, P Kristensen, R M Brosh, D K Orren, V A Bohr, B F Clark

8 Citationer (Scopus)

Abstract

A procedure was established for selecting phage antibodies (phage-abs) from phage-displayed antibody repertoires by panning against proteins, separated by sodium dodecyl phosphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose membranes (Western blots). This immobilization strategy is applicable for secondary rounds of panning in selections against semipurified proteins, and directs the selection toward antibodies suitable as immunochemical reagents in Western blots. In model experiments, enrichment factors as high as 1.9x10(5) were obtained in a single round of panning. Furthermore, we demonstrate the application of this approach by selection of phage-abs recognizing the human Werner protein, which is defective in a premature aging syndrome.

Originalsprog	Engelsk
Tidsskrift	Electrophoresis
Vol/bind	21
Udgave nummer	3
Sider (fra-til)	509-16
Antal sider	8
ISSN	0173-0835
DOI	https://doi.org/10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.0.CO;2-5
Status	Udgivet - feb. 2000
Udgivet eksternt	Ja

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10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.0.CO;2-5

Citationsformater

@article{e5e654c7c2ec4cdc8975e0750e341c58,

title = "Identification of phage antibodies toward the Werner protein by selection on Western blots",

abstract = "A procedure was established for selecting phage antibodies (phage-abs) from phage-displayed antibody repertoires by panning against proteins, separated by sodium dodecyl phosphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose membranes (Western blots). This immobilization strategy is applicable for secondary rounds of panning in selections against semipurified proteins, and directs the selection toward antibodies suitable as immunochemical reagents in Western blots. In model experiments, enrichment factors as high as 1.9x10(5) were obtained in a single round of panning. Furthermore, we demonstrate the application of this approach by selection of phage-abs recognizing the human Werner protein, which is defective in a premature aging syndrome.",

keywords = "Antibodies, Viral/immunology, Bacteriophages/immunology, Base Sequence, Blotting, Western, DNA Fingerprinting, DNA Helicases/immunology, DNA Primers, Enzyme-Linked Immunosorbent Assay, Exodeoxyribonucleases, Humans, Polymerase Chain Reaction, RecQ Helicases, Werner Syndrome Helicase",

author = "Peter Ravn and Svend Kjaer and Jensen, {Kristian Hobolt} and T Wind and Jensen, {K B} and P Kristensen and Brosh, {R M} and Orren, {D K} and Bohr, {V A} and Clark, {B F}",

year = "2000",

month = feb,

doi = "10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.0.CO;2-5",

language = "English",

volume = "21",

pages = "509--16",

journal = "Electrophoresis",

issn = "0173-0835",

publisher = "Wiley - V C H Verlag GmbH & Co. KGaA",

number = "3",

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TY - JOUR

T1 - Identification of phage antibodies toward the Werner protein by selection on Western blots

AU - Ravn, Peter

AU - Kjaer, Svend

AU - Jensen, Kristian Hobolt

AU - Wind, T

AU - Jensen, K B

AU - Kristensen, P

AU - Brosh, R M

AU - Orren, D K

AU - Bohr, V A

AU - Clark, B F

PY - 2000/2

Y1 - 2000/2

N2 - A procedure was established for selecting phage antibodies (phage-abs) from phage-displayed antibody repertoires by panning against proteins, separated by sodium dodecyl phosphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose membranes (Western blots). This immobilization strategy is applicable for secondary rounds of panning in selections against semipurified proteins, and directs the selection toward antibodies suitable as immunochemical reagents in Western blots. In model experiments, enrichment factors as high as 1.9x10(5) were obtained in a single round of panning. Furthermore, we demonstrate the application of this approach by selection of phage-abs recognizing the human Werner protein, which is defective in a premature aging syndrome.

AB - A procedure was established for selecting phage antibodies (phage-abs) from phage-displayed antibody repertoires by panning against proteins, separated by sodium dodecyl phosphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose membranes (Western blots). This immobilization strategy is applicable for secondary rounds of panning in selections against semipurified proteins, and directs the selection toward antibodies suitable as immunochemical reagents in Western blots. In model experiments, enrichment factors as high as 1.9x10(5) were obtained in a single round of panning. Furthermore, we demonstrate the application of this approach by selection of phage-abs recognizing the human Werner protein, which is defective in a premature aging syndrome.

KW - Antibodies, Viral/immunology

KW - Bacteriophages/immunology

KW - Base Sequence

KW - Blotting, Western

KW - DNA Fingerprinting

KW - DNA Helicases/immunology

KW - DNA Primers

KW - Enzyme-Linked Immunosorbent Assay

KW - Exodeoxyribonucleases

KW - Humans

KW - Polymerase Chain Reaction

KW - RecQ Helicases

KW - Werner Syndrome Helicase

U2 - 10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.0.CO;2-5

DO - 10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.0.CO;2-5

M3 - Journal article

C2 - 10726750

SN - 0173-0835

VL - 21

SP - 509

EP - 516

JO - Electrophoresis

JF - Electrophoresis

IS - 3

ER -

Identification of phage antibodies toward the Werner protein by selection on Western blots

Abstract

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