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Mapping of lanthanide-tagged proteins on western blot paper using microextraction-inductively coupled plasma-mass spectrometry

Journal Article · · Journal of Analytical Atomic Spectrometry
DOI:https://doi.org/10.1039/D4JA00440J· OSTI ID:2549507
 [1];  [2];  [3];  [1];  [1];  [1];  [1];  [1]
  1. Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
  2. Berry College, Mt Berry, GA (United States)
  3. Univ. of Tennessee, Knoxville, TN (United States). Oak Ridge Innovation Institute
+ Show Author Affiliations
A microextraction (ME) sampling system, paired with inductively coupled plasma-mass spectrometry (ICP-MS), was employed to spatially analyze proteins tagged with lanthanum (La), gadolinium (Gd), or terbium (Tb) on the surface of western blot paper. The proteins were covalently tagged, separated via gel electrophoresis, and transferred to western blot paper for analysis by ME-ICP-MS. The ME-ICP-MS method enables the direct sampling of the tagged species on the western blot paper, without any sample preparation. Traditionally, the tagged analyte would need to be stained, excised, and digested to be analyzed by ICP-MS for its elemental and isotopic characterization. Preliminary detection limits for the ME-ICP-MS method applied to western blot paper were established to be 564, 54, and 2.5 fg for La, Gd, and Tb, respectively. The developed ME-ICP-MS method was compared to laser ablation (LA) ICP-MS, another direct solid sampling technique; it was readily determined that ME-ICP-MS can effectively map the elemental constituents on the western blot paper with comparable analysis time and measurement sensitivity. The analysis time per 2 × 4 mm extraction is ∼1 minute; if protein spots are directly targeted (rather than systematically mapping the entire blot paper), the analysis time per protein spot is ∼1 min. This developed method proved to be fast, effective, and accessible for correlating protein molecular weight with the detection of the inorganic tagant. The ME-ICP-MS approach could be widely applicable in research areas that involve metal-tagged protein bioconjugates, such as in the development of diagnostic and therapeutic agents and other biochemical probes.
Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE; USDOE National Nuclear Security Administration (NNSA); USDOE Office of Science (SC)
Grant/Contract Number:
AC05-00OR22725
OSTI ID:
2549507
Alternate ID(s):
OSTI ID: 2573445
Journal Information:
Journal of Analytical Atomic Spectrometry, Journal Name: Journal of Analytical Atomic Spectrometry Journal Issue: 4 Vol. 40; ISSN 0267-9477; ISSN 1364-5544
Publisher:
Royal Society of ChemistryCopyright Statement
Country of Publication:
United States
Language:
English

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