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Title of article :
Selective enrichment of monospecific polyclonal antibodies for antibody-based proteomics efforts
Author/Authors :
Agaton، نويسنده , , Charlotta and Falk، نويسنده , , Ronny and Hِidén Guthenberg، نويسنده , , Ingmarie and Gِstring، نويسنده , , Lovisa and Uhlén، نويسنده , , Mathias and Hober، نويسنده , , Sophia، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2004
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Abstract :
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.
Keywords :
PROTEOMICS , Immobilized metal-ion affinity chromatography , affinity chromatography , antibodies , Protein epitope signature tags
Journal title :
Journal of Chromatography A
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Link To Document :