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

Methods Mol Biol. 2009:483:313-24. doi: 10.1007/978-1-59745-407-0_18.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Molecular Weight
  • Plant Extracts / chemistry*
  • Plants, Genetically Modified
  • Recombinant Proteins / analysis
  • Recombinant Proteins / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Plant Extracts
  • Recombinant Proteins