On-chip detection of low-molecular-weight recombinant proteins in plant crude extracts by SELDI-TOF MS.
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.
Badri
Amine M
AM
Département de phytologie, Université Laval, Québec, Canada.
Coenen
Karine
K
Vaillancourt
Louis-Philippe
LP
Goulet
Charles
C
Michaud
Dominique
D
D008970:Molecular Weight; D010936:Plant Extracts / Q000737:chemistry*; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D010936:Plant Extracts; D011994:Recombinant Proteins