Phage display derived human monoclonal antibodies isolated by binding to the surface of live primary breast cancer cells recognize GRP78

Charlotte G Jakobsen, Nicolaj Rasmussen, Anne-Vibeke Laenkholm, Henrik Ditzel

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

 
Udgivelsesdato: 2007-Oct-1
OriginalsprogEngelsk
TidsskriftCancer Research
Vol/bind67
Udgave nummer19
Sider (fra-til)9507-17
Antal sider10
ISSN0008-5472
DOI
StatusUdgivet - 1. okt. 2007

Fingeraftryk

Neoplasms
Libraries
Melanoma
Membrane Proteins
Clinical Trials
Cell Line

Citer dette

Jakobsen, Charlotte G ; Rasmussen, Nicolaj ; Laenkholm, Anne-Vibeke ; Ditzel, Henrik. / Phage display derived human monoclonal antibodies isolated by binding to the surface of live primary breast cancer cells recognize GRP78. I: Cancer Research. 2007 ; Bind 67, Nr. 19. s. 9507-17.
@article{f7e6cd80cb5111dc8674000ea68e967b,
title = "Phage display derived human monoclonal antibodies isolated by binding to the surface of live primary breast cancer cells recognize GRP78",
abstract = "Clinical trials using monoclonal antibodies (mAb) against cell-surface markers have yielded encouraging therapeutic results in several cancer types. Generally, however, anticancer antibodies are only efficient against a subpopulation of cancers, and there is a strong need for identification of novel targets and human antibodies against them. We have isolated single-chain human mAbs from a large na{\"i}ve antibody phage display library by panning on a single-cell suspension of freshly isolated live cancer cells from a human breast cancer specimen, and these antibodies were shown to specifically recognize cancer-associated cell-surface proteins. One of the isolated human antibody fragments, Ab39, recognizes a cell-surface antigen expressed on a subpopulation of cancer cell lines of different origins. Immunohistochemical analysis of a large panel of cancerous and normal tissues showed that Ab39 bound strongly to several cancers, including 45{\%} breast carcinomas, 35{\%} lung cancers, and 86{\%} melanomas, but showed no or weak binding to normal tissues. A yeast two-hybrid screen of a large human testis cDNA library identified the glucose-regulated protein of 78 kDa (GRP78) as the antigen recognized by Ab39. The interaction was confirmed by colocalization studies and antibody competition experiments that also mapped the epitope recognized by Ab39 to the COOH terminus of GRP78. The expression of GRP78 on the surface of cancer cells, but not normal cells, makes it an attractive target for cancer therapies including mAb-based immunotherapy. Our results suggest that the human antibody Ab39 may be a useful starting point for further genetic optimization that could render it a useful diagnostic and therapeutic reagent for a variety of cancers",
keywords = "Aged, 80 and over, Antibodies, Monoclonal, Antigens, Neoplasm, Blotting, Western, Breast Neoplasms, Carcinoma, Ductal, Cell Line, Tumor, Epitope Mapping, Female, Heat-Shock Proteins, Humans, Immunoglobulin Fragments, Immunohistochemistry, Molecular Chaperones, Peptide Library",
author = "Jakobsen, {Charlotte G} and Nicolaj Rasmussen and Anne-Vibeke Laenkholm and Henrik Ditzel",
year = "2007",
month = "10",
day = "1",
doi = "10.1158/0008-5472.CAN-06-4686",
language = "English",
volume = "67",
pages = "9507--17",
journal = "Cancer Research",
issn = "0008-5472",
publisher = "American Association for Cancer Research (A A C R)",
number = "19",

}

Phage display derived human monoclonal antibodies isolated by binding to the surface of live primary breast cancer cells recognize GRP78. / Jakobsen, Charlotte G; Rasmussen, Nicolaj; Laenkholm, Anne-Vibeke; Ditzel, Henrik.

I: Cancer Research, Bind 67, Nr. 19, 01.10.2007, s. 9507-17.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Phage display derived human monoclonal antibodies isolated by binding to the surface of live primary breast cancer cells recognize GRP78

AU - Jakobsen, Charlotte G

AU - Rasmussen, Nicolaj

AU - Laenkholm, Anne-Vibeke

AU - Ditzel, Henrik

PY - 2007/10/1

Y1 - 2007/10/1

N2 - Clinical trials using monoclonal antibodies (mAb) against cell-surface markers have yielded encouraging therapeutic results in several cancer types. Generally, however, anticancer antibodies are only efficient against a subpopulation of cancers, and there is a strong need for identification of novel targets and human antibodies against them. We have isolated single-chain human mAbs from a large naïve antibody phage display library by panning on a single-cell suspension of freshly isolated live cancer cells from a human breast cancer specimen, and these antibodies were shown to specifically recognize cancer-associated cell-surface proteins. One of the isolated human antibody fragments, Ab39, recognizes a cell-surface antigen expressed on a subpopulation of cancer cell lines of different origins. Immunohistochemical analysis of a large panel of cancerous and normal tissues showed that Ab39 bound strongly to several cancers, including 45% breast carcinomas, 35% lung cancers, and 86% melanomas, but showed no or weak binding to normal tissues. A yeast two-hybrid screen of a large human testis cDNA library identified the glucose-regulated protein of 78 kDa (GRP78) as the antigen recognized by Ab39. The interaction was confirmed by colocalization studies and antibody competition experiments that also mapped the epitope recognized by Ab39 to the COOH terminus of GRP78. The expression of GRP78 on the surface of cancer cells, but not normal cells, makes it an attractive target for cancer therapies including mAb-based immunotherapy. Our results suggest that the human antibody Ab39 may be a useful starting point for further genetic optimization that could render it a useful diagnostic and therapeutic reagent for a variety of cancers

AB - Clinical trials using monoclonal antibodies (mAb) against cell-surface markers have yielded encouraging therapeutic results in several cancer types. Generally, however, anticancer antibodies are only efficient against a subpopulation of cancers, and there is a strong need for identification of novel targets and human antibodies against them. We have isolated single-chain human mAbs from a large naïve antibody phage display library by panning on a single-cell suspension of freshly isolated live cancer cells from a human breast cancer specimen, and these antibodies were shown to specifically recognize cancer-associated cell-surface proteins. One of the isolated human antibody fragments, Ab39, recognizes a cell-surface antigen expressed on a subpopulation of cancer cell lines of different origins. Immunohistochemical analysis of a large panel of cancerous and normal tissues showed that Ab39 bound strongly to several cancers, including 45% breast carcinomas, 35% lung cancers, and 86% melanomas, but showed no or weak binding to normal tissues. A yeast two-hybrid screen of a large human testis cDNA library identified the glucose-regulated protein of 78 kDa (GRP78) as the antigen recognized by Ab39. The interaction was confirmed by colocalization studies and antibody competition experiments that also mapped the epitope recognized by Ab39 to the COOH terminus of GRP78. The expression of GRP78 on the surface of cancer cells, but not normal cells, makes it an attractive target for cancer therapies including mAb-based immunotherapy. Our results suggest that the human antibody Ab39 may be a useful starting point for further genetic optimization that could render it a useful diagnostic and therapeutic reagent for a variety of cancers

KW - Aged, 80 and over

KW - Antibodies, Monoclonal

KW - Antigens, Neoplasm

KW - Blotting, Western

KW - Breast Neoplasms

KW - Carcinoma, Ductal

KW - Cell Line, Tumor

KW - Epitope Mapping

KW - Female

KW - Heat-Shock Proteins

KW - Humans

KW - Immunoglobulin Fragments

KW - Immunohistochemistry

KW - Molecular Chaperones

KW - Peptide Library

U2 - 10.1158/0008-5472.CAN-06-4686

DO - 10.1158/0008-5472.CAN-06-4686

M3 - Journal article

VL - 67

SP - 9507

EP - 9517

JO - Cancer Research

JF - Cancer Research

SN - 0008-5472

IS - 19

ER -