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A3 -- a novel colon and pancreatic cancer reactive antibody from a primate phage library selected using intact tumour cells

Tordsson, Jesper LU ; Lavasani, Shahram LU ; Ohlsson, Lennart; Chassale, Pia LU ; Svedberg, Helena LU ; Abrahmsen, Lars and Brodin, Thomas (2000) In International Journal of Cancer 87(4). p.559-568
Abstract
The identification of novel tumour-associated antigens (TAAs) is pivotal for progression in the fields of tumour immunotherapy and diagnosis. In the present study, we have developed, based on flow cytometric evaluation and use of a mini-library composed of specific antibody clones linked to different antibiotic resistance markers, methods for positive and subtractive selection of phage antibodies employing intact cells as the antigen source. An scFv phage library (2.7 x 10(7)) was constructed from a primate (Macaca fascicularis) immunised with pooled human colon carcinomas. This library was selected for 3 rounds by binding to Colo 205 colon adenocarcinoma cells and proteolytic elution followed by phage amplification. Several antibodies... (More)
The identification of novel tumour-associated antigens (TAAs) is pivotal for progression in the fields of tumour immunotherapy and diagnosis. In the present study, we have developed, based on flow cytometric evaluation and use of a mini-library composed of specific antibody clones linked to different antibiotic resistance markers, methods for positive and subtractive selection of phage antibodies employing intact cells as the antigen source. An scFv phage library (2.7 x 10(7)) was constructed from a primate (Macaca fascicularis) immunised with pooled human colon carcinomas. This library was selected for 3 rounds by binding to Colo 205 colon adenocarcinoma cells and proteolytic elution followed by phage amplification. Several antibodies reactive with colon carcinomas and with restricted reactivity to a few epithelial normal tissues were identified by immunohistochemistry. One clone, A3 scFv, recognised an epitope that was homogeneously expressed in 11/11 of colon and 4/4 pancreatic carcinomas studied and in normal tissue restricted to subtypes of epithelia in the gastrointestinal tract. The A3 scFv had an apparent overall affinity approximately 100-fold higher than an A3 Fab, suggesting binding of scFv homodimers. The cell surface density of the A3 epitope, calculated on the basis of Fab binding, was exceptionally high, approaching 3 million per cell. We also demonstrate efficient T-cell-mediated killing of colon cancer cells coated with A3 scFv fused to the low MHC class II binding superantigen mutant SEA(D227A). The identified A3 molecule thus represents a TAA with properties that suggest its use for immunotherapy of colon and pancreatic cancer. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
International Journal of Cancer
volume
87
issue
4
pages
559 - 568
publisher
John Wiley & Sons
external identifiers
  • pmid:10918198
  • scopus:0033897739
ISSN
0020-7136
DOI
10.1002/1097-0215(20000815)87:4<559::AID-IJC16>3.0.CO;2-#
language
English
LU publication?
yes
id
0e237c4f-57e1-403a-bf76-327f13a9e6f6 (old id 1117075)
date added to LUP
2008-07-02 16:13:02
date last changed
2017-01-01 05:19:21
@article{0e237c4f-57e1-403a-bf76-327f13a9e6f6,
  abstract     = {The identification of novel tumour-associated antigens (TAAs) is pivotal for progression in the fields of tumour immunotherapy and diagnosis. In the present study, we have developed, based on flow cytometric evaluation and use of a mini-library composed of specific antibody clones linked to different antibiotic resistance markers, methods for positive and subtractive selection of phage antibodies employing intact cells as the antigen source. An scFv phage library (2.7 x 10(7)) was constructed from a primate (Macaca fascicularis) immunised with pooled human colon carcinomas. This library was selected for 3 rounds by binding to Colo 205 colon adenocarcinoma cells and proteolytic elution followed by phage amplification. Several antibodies reactive with colon carcinomas and with restricted reactivity to a few epithelial normal tissues were identified by immunohistochemistry. One clone, A3 scFv, recognised an epitope that was homogeneously expressed in 11/11 of colon and 4/4 pancreatic carcinomas studied and in normal tissue restricted to subtypes of epithelia in the gastrointestinal tract. The A3 scFv had an apparent overall affinity approximately 100-fold higher than an A3 Fab, suggesting binding of scFv homodimers. The cell surface density of the A3 epitope, calculated on the basis of Fab binding, was exceptionally high, approaching 3 million per cell. We also demonstrate efficient T-cell-mediated killing of colon cancer cells coated with A3 scFv fused to the low MHC class II binding superantigen mutant SEA(D227A). The identified A3 molecule thus represents a TAA with properties that suggest its use for immunotherapy of colon and pancreatic cancer.},
  author       = {Tordsson, Jesper and Lavasani, Shahram and Ohlsson, Lennart and Chassale, Pia and Svedberg, Helena and Abrahmsen, Lars and Brodin, Thomas},
  issn         = {0020-7136},
  language     = {eng},
  number       = {4},
  pages        = {559--568},
  publisher    = {John Wiley & Sons},
  series       = {International Journal of Cancer},
  title        = {A3 -- a novel colon and pancreatic cancer reactive antibody from a primate phage library selected using intact tumour cells},
  url          = {http://dx.doi.org/10.1002/1097-0215(20000815)87:4<559::AID-IJC16>3.0.CO;2-#},
  volume       = {87},
  year         = {2000},
}