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Characterization of...
Characterization of PrEST-based antibodies towards human Cytokeratin-17
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- Larsson, Karin (författare)
- KTH,Proteomik
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- Eriksson, Cecilia (författare)
- KTH,Proteomik
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- Schwenk, Jochen. M. (författare)
- KTH,Proteomik
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- Berglund, Lisa (författare)
- KTH,Proteomik
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- Wester, Kenneth (författare)
- Uppsala universitet,Institutionen för genetik och patologi
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- Uhlén, Mattias (författare)
- KTH,Proteomik
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- Hober, Sophia (författare)
- KTH,Proteomik
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Wernérus, H. (författare)
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(creator_code:org_t)
- Elsevier BV, 2009
- 2009
- Engelska.
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Ingår i: JIM - Journal of Immunological Methods. - : Elsevier BV. - 0022-1759 .- 1872-7905. ; 342:1-2, s. 20-32
- Relaterad länk:
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http://www.sciencedi...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Antibody-based proteomics efforts depend on validated antibodies to ensure correct annotation of analyzed proteins. We have previously argued that a low sequence identity to other proteins is a key feature for antigens used in antibody generation. Thus, a major challenge for whole-proteome studies is how to address families of highly sequence related proteins within the context of generating specific antibodies. In this study, two non-overlapping parts of human Cytokeratin-17, a protein belonging to the intermediate filament family of highly sequence-related proteins, were selected as a model system to study the specificity and cross reactivity of antibodies generated towards such a target. These recombinantly produced Protein Epitope Signature Tags (PrESTs) were immunized in five rabbits each and the batch-to-batch variations in the obtained immune responses were studied by mapping of linear epitopes using synthetic overlapping peptides. The obtained results showed a similar but not identical immune response in the respective antibody groups with a limited number of epitopes being identified. Immunohistochemical analysis of the affinity purified monospecific antibodies on tissue micro arrays resulted in a general recognition of human cytokeratins for all analyzed binders whereas antibodies identified as binding to the most unique parts of the PrESTs showed the most Cytokeratin-17 like staining. The data presented here support the strategy to use sequence identity scores as the main criteria for antigen selection but also indicate the possibility to instead produce a single antibody recognizing a defined group of proteins when the intended targets overall sequence identity score is too high. This type of group-specific antibodies would be an important tool for antibody-based projects aiming for a complete coverage of the human proteome.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
Nyckelord
- Cytokeratin-17
- PrEST
- Epitope mapping
- Antibody
- MEDICINE
- MEDICIN
- Bioengineering
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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