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Expression of Prostate Specific Antigen on the Surface of a Filamentous Phage

Eerola, Riitta ; Saviranta, Petri ; Lilja, Hans LU orcid ; Pettersson, Kim and Lövgren, Timo (1994) In Biochemical and Biophysical Research Communications 200(3). p.1346-1352
Abstract

We have constructed two phagemid vectors containing the gene coding for human Prostate Specific Antigen (PSA) translationally fused to a pelB signal sequence and minor coat protein III of phage fd leading to phage particles that carry PSA on their tips. Phages were characterized by Western blotting and by panning, using biotinylated monoclonal anti-PSA antibodies immobilized onto streptavidin-coated microtitration wells. Binding of PSA-phages was 102- to 5×103-fold more effective than that of wildtype phages. Trypsin treatment of the phage abolished the specific binding. Competitive panning with different concentrations of added purified PSA demonstrated that binding was PSA specific. Finally, Western-blot analysis... (More)

We have constructed two phagemid vectors containing the gene coding for human Prostate Specific Antigen (PSA) translationally fused to a pelB signal sequence and minor coat protein III of phage fd leading to phage particles that carry PSA on their tips. Phages were characterized by Western blotting and by panning, using biotinylated monoclonal anti-PSA antibodies immobilized onto streptavidin-coated microtitration wells. Binding of PSA-phages was 102- to 5×103-fold more effective than that of wildtype phages. Trypsin treatment of the phage abolished the specific binding. Competitive panning with different concentrations of added purified PSA demonstrated that binding was PSA specific. Finally, Western-blot analysis confirmed that full-length and shorter degradation products of the fusion between PSA and protein III were visible as probed with anti-PSA antibody.

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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
in
Biochemical and Biophysical Research Communications
volume
200
issue
3
pages
1346 - 1352
publisher
Elsevier
external identifiers
  • scopus:0028275233
  • pmid:7514400
ISSN
0006-291X
DOI
10.1006/bbrc.1994.1599
language
English
LU publication?
yes
id
ec4adc11-158c-49ed-9232-57d5e94dfb48
date added to LUP
2022-12-08 12:58:15
date last changed
2024-01-02 13:24:44
@article{ec4adc11-158c-49ed-9232-57d5e94dfb48,
  abstract     = {{<p>We have constructed two phagemid vectors containing the gene coding for human Prostate Specific Antigen (PSA) translationally fused to a pelB signal sequence and minor coat protein III of phage fd leading to phage particles that carry PSA on their tips. Phages were characterized by Western blotting and by panning, using biotinylated monoclonal anti-PSA antibodies immobilized onto streptavidin-coated microtitration wells. Binding of PSA-phages was 10<sup>2</sup>- to 5×10<sup>3</sup>-fold more effective than that of wildtype phages. Trypsin treatment of the phage abolished the specific binding. Competitive panning with different concentrations of added purified PSA demonstrated that binding was PSA specific. Finally, Western-blot analysis confirmed that full-length and shorter degradation products of the fusion between PSA and protein III were visible as probed with anti-PSA antibody.</p>}},
  author       = {{Eerola, Riitta and Saviranta, Petri and Lilja, Hans and Pettersson, Kim and Lövgren, Timo}},
  issn         = {{0006-291X}},
  language     = {{eng}},
  month        = {{05}},
  number       = {{3}},
  pages        = {{1346--1352}},
  publisher    = {{Elsevier}},
  series       = {{Biochemical and Biophysical Research Communications}},
  title        = {{Expression of Prostate Specific Antigen on the Surface of a Filamentous Phage}},
  url          = {{http://dx.doi.org/10.1006/bbrc.1994.1599}},
  doi          = {{10.1006/bbrc.1994.1599}},
  volume       = {{200}},
  year         = {{1994}},
}