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On-chip detection of low-molecular-weight recombinant proteins in plant crude extracts by SELDI-TOF MS.

Badri, Amine M; Coenen, Karine; Vaillancourt, Louis-Philippe; Goulet, Charles; Michaud, Dominique.
Methods Mol Biol ; 483: 313-24, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19183907
This chapter presents a general procedure for the on-chip detection and quantitation of low-molecular-weight recombinant proteins in transgenic plant crude extracts by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). A protocol is first described to detect the protein of interest in crude protein extracts of transgenic plant lines, by differential protein mapping against similar extracts from a control, nontransgenic line. A complementary protocol is then presented to generate a standard curve with the SELDI system, allowing the protein to be quantified in different transgenic lines. Overall, this procedure may be carried out within a few hours, without the need for prior purification or enrichment of the recombinant protein.