Identification of phage antibodies toward the Werner protein by selection on Western blots
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Identification of phage antibodies toward the Werner protein by selection on Western blots. / Ravn, Peter; Kjaer, Svend; Jensen, Kristian Hobolt; Wind, T; Jensen, K B; Kristensen, P; Brosh, R M; Orren, D K; Bohr, V A; Clark, B F.
In: Electrophoresis, Vol. 21, No. 3, 02.2000, p. 509-16.Research output: Contribution to journal › Journal article › Research › peer-review
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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
VL - 21
SP - 509
EP - 516
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 3
ER -
ID: 200573317