Change search
ReferencesLink to record
Permanent link

Direct link
Generation of HER2 monoclonal antibodies using epitopes of a rabbit polyclonal antibody
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-8993-048X
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
2014 (English)In: New Biotechnology, ISSN 1871-6784, Vol. 31, no 1, 35-43 p.Article in journal (Refereed) Published
Abstract [en]

One of the issues in using polyclonal antibodies is the limited amount of reagent available from an immunisation, leading to batch-to-batch variation and difficulties in obtaining the same antibody performance when the same antigen is re-immunised into several separate animals. This led to the development of hybridoma technology allowing, at least theoretically, for an unlimited production of a specific binder. Nevertheless, polyclonal antibodies are widely used in research and diagnostics and there exists a need for robust methods to convert a polyclonal antibody with good binding performance into a renewable monoclonal with identical or similar binding specificity. Here we have used precise information regarding the functional recognition sequence (epitope) of a rabbit polyclonal antibody with attractive binding characteristics as the basis for generation of a renewable mouse monoclonal antibody. First, the original protein fragment antigen was used for immunisation and generation of mouse hybridoma, without obtaining binders to the same epitope region. Instead a peptide designed using the functional epitope and structural information was synthesised and used for hybridoma production. Several of the monoclonal antibodies generated were found to have similar binding characteristics to those of the original polyclonal antibody. These monoclonal antibodies detected native HER2 on cell lines and were also able to stain HER2 in immunohistochemistry using xenografted mice, as well as human normal and cancer tissues.

Place, publisher, year, edition, pages
2014. Vol. 31, no 1, 35-43 p.
Keyword [en]
Batch-to-batch variations, Binding characteristics, Binding specificities, Hybridoma technology, Immunohistochemistry, Polyclonal antibody, Recognition sequence, Structural information
National Category
Biochemistry and Molecular Biology
URN: urn:nbn:se:kth:diva-139254DOI: 10.1016/j.nbt.2013.10.002ISI: 000328131200004ScopusID: 2-s2.0-84889568450OAI: diva2:685945
EU, FP7, Seventh Framework Programme, 241481VinnovaKnut and Alice Wallenberg FoundationScience for Life Laboratory - a national resource center for high-throughput molecular bioscience

QC 20140110

Available from: 2014-01-10 Created: 2014-01-08 Last updated: 2014-01-10Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textScopus

Search in DiVA

By author/editor
Hu, Francis JingxinUhlén, MathiasRockberg, Johan
By organisation
Proteomics and NanobiotechnologyScience for Life Laboratory, SciLifeLab
In the same journal
New Biotechnology
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 30 hits
ReferencesLink to record
Permanent link

Direct link