Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Discovery of epitopes for targeting the human epidermal growth factor receptor 2 (HER2) with antibodies
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0002-9977-5724
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0001-8141-8449
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0001-8993-048X
2009 (English)In: Molecular Oncology, ISSN 1574-7891, Vol. 3, no 3, 238-247 p.Article in journal (Refereed) Published
Abstract [en]

Antibodies have become valuable therapeutic agents for targeting of extracellular proteins in various diseases, including cancer, autoimmunity and cardiovascular disorders. For breast cancer, antibodies targeting the human HER2 have been shown to result in cell growth inhibition both in vitro and in patients with breast tumors. There is evidence to suggest that targeting multiple HER2 epitopes may result in increased growth inhibition making it interesting to find antibodies targeting new epitopes. Here, we report on a new scheme to discover antibodies directed to new epitopes using the extracellular domain of the HER2 as a model. Polyclonal antibodies were generated using recombinant protein fragments and affinity purified fractions of the antibodies were functionally characterized and precisely epitope mapped using bacterial surface display. Polyclonal antibodies towards a 127 amino acid recombinant protein fragment spanning between domains II and III of the HER2 were shown to bind to human ductal carcinoma cell line BT474 resulting in growth inhibition. Affinity purification demonstrated that antibodies to two separate regions from the N- and C-terminal end of the fragment exhibited the growth inhibition. Epitope mapping of the C-terminal antibodies revealed a 25 amino acid region (LPESFDGDPASNTAPLQPEQLQVF) with two distinct epitopes mediating efficient growth inhibition. The results suggest that antibodies directed towards this region of domain III of the HER2, distinct from the well-known monoclonal antibodies trastuzumab and pertuzumab, bind to the HER2 on living cells and exhibit growth inhibition. The work describes a new strategy to develop antibodies directed to non-overlapping epitopes and shows a path of pursuit to explore the epitope space of a target protein.

Place, publisher, year, edition, pages
2009. Vol. 3, no 3, 238-247 p.
Keyword [en]
Antibody, Growth inhibition, HER2, Epitope mapping, human-breast-cancer, polyclonal antibodies, therapy, trastuzumab, proteomics, pertuzumab, generation, herceptin, survival, disease
National Category
Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-18497DOI: 10.1016/j.molonc.2009.01.003ISI: 000266853200006Scopus ID: 2-s2.0-67349155587OAI: oai:DiVA.org:kth-18497DiVA: diva2:336544
Note
QC 20100525. Tidigare titel: Targeting the human epidermal growth factor receptor 2 (HER2) with antibodiesAvailable from: 2010-08-05 Created: 2010-08-05 Last updated: 2011-01-14Bibliographically approved
In thesis
1. Methods for Generation and Characterization of Monospecific Antibodies
Open this publication in new window or tab >>Methods for Generation and Characterization of Monospecific Antibodies
2008 (English)Doctoral thesis, comprehensive summary (Other scientific)
Abstract [en]

Recent advances in biotechnology have generated possibilities to investigate and measure parts of life previously left for believers to explain. Utilizing the same book of recipes, the genome, our cells produce selections of proteins at a time and thereby niche into a multitude of specialized cell types, tissues and organs comprising our body. Knowledge of the precise protein composition in a given organ at normal and disease condition would be of invaluable importance, both for identification of disease causes and the design of new pharmaceuticals, as well as for a deeper understanding of the processes of life. This doctoral thesis describes the start and progress of a visionary project (HPR) to localize all human proteins in our body, with emphasis on the generation and characterization of antibodies used as protein targeting missiles. To facilitate the identification of one human protein in a complex environment like our body, it is of significant importance to have precise and specific means of detection. The first two papers (I-II), describe software developed for generation of monospecific antibodies satisfying such needs, using a set of rules for antigen optimization. Five years after project start a large amount of antibodies with documented characteristics have been generated. The third paper (III), illustrates an attempt to sieve these antibody characteristics to develop a tool, for further improvement of antigen selection, based on the correlation between antigen sequence and amount of specific antibody generated.Having a panel of protein-specific antibodies is a possession of a great value, not only for localization studies, but also as possible target-directed pharmaceuticals. In such cases, knowledge of the precise epitope recognized by the antibody on its target protein, is an important aid, both for understanding its effect as well as unwanted cross-reactivity. Paper (IV) describes the development of a high-resolution method for epitope mapping of antibodies using staphylococcal display. An application of the method is described in the last paper (V) where it is used to map an anti-HER2 monospecific antibody with growth-inhibiting effects on breast cancer cells. The monospecific antibody was fractionated into separate populations and five novel epitopes related to cancer cell growth-inhibition was determined.Altogether these methods are valuable tools for generation and characterization of monospecific antibodies.

Place, publisher, year, edition, pages
Stockholm: KTH, 2008. xii, 66 p.
Series
Trita-BIO-Report, ISSN 1654-2312 ; 2008:20
Keyword
antibody, epitope mapping, HER2, human protein atlas, immunogenicity, proteomics
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-9338 (URN)978-91-7415-139-8 (ISBN)
Public defence
2008-10-31, F3, KTH, Lindstedtsvägen 26, Stockholm, 10:00 (English)
Opponent
Supervisors
Note
QC 20100907Available from: 2008-10-21 Created: 2008-10-21 Last updated: 2010-09-07Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textScopus

Authority records BETA

Schwenk, Jochen M.Uhlén, Mathias

Search in DiVA

By author/editor
Rockberg, JohanSchwenk, Jochen M.Uhlén, Mathias
By organisation
Proteomics
Industrial Biotechnology

Search outside of DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 133 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf