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Selective enrichmen...
Selective enrichment of monospecific polyclonal antibodies for antibody-based proteomics efforts
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- Agaton, Charlotta (author)
- KTH,Bioteknologi
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- Falk, Ronny (author)
- KTH,Bioteknologi
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- Uhlén, Mathias (author)
- KTH,Bioteknologi
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- Hober, Sophia (author)
- KTH,Bioteknologi
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(creator_code:org_t)
- Elsevier BV, 2004
- 2004
- English.
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In: Journal of Chromatography A. - : Elsevier BV. - 0021-9673 .- 1873-3778. ; 1043, s. 33-40
- Related links:
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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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Abstract
Subject headings
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- A high stringency protocol, suitable for systematic purification of polyclonal antibodies, is described. The procedure is designed to allow the generation of target protein-specific antibodies suitable for functional annotation of proteins. Antibodies were generated by immunization with recombinantly produced affinity-tagged target proteins. To obtain stringent recovery of the antibodies, a two-step affinity chromatography principle was devised to first deplete the affinity tag-specific antibodies followed by a second step for affinity capture of the target protein-specific antibodies. An analytical dot-blot array system was developed to analyze the cross-reactivity of the affinity-purified antibodies. The results suggest that the protocol can be used in a highly parallel and automated manner to generate mono-specific polyclonal antibodies for large-scale, antibody-based proteomics efforts, i.e. affinity proteomics.
Subject headings
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
Keyword
- Affinity chromatography; Antibodies; Immobilized metal-ion affinity chromatography; Protein epitope signature tags; Proteomics
- Bioengineering
- Bioteknik
Publication and Content Type
- ref (subject category)
- art (subject category)
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