14C radiolabeling of proteins to monitor biodistribution of ingested proteins

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DOIResolve DOI: http://doi.org/10.1016/j.ab.2010.11.028
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TypeArticle
Journal titleAnalytical Biochemistry
ISSN0003-2697
Volume410
Issue1
Pages5761; # of pages: 5
Subjectbovine serum albumin; casein; CD14 antigen; methyl iodide; methyl iodide carbon 14; unclassified drug; animal experiment; animal tissue; article; drug accumulation; drug distribution; drug efficacy; isotope labeling; nonhuman; priority journal; radioactivity; rat; Animals; Carbon Radioisotopes; Cattle; Eating; Freeze Drying; Humans; Isotope Labeling; Methylation; Proteins; Rats; Rats, Sprague-Dawley; Solubility; Bacteria (microorganisms); Bovinae; Rattus
AbstractThe economical preparation of microgram quantities of 14C- labeled proteins by in vacuo methylation with methyl iodide is described. The 14C radiolabeling was achieved by the covalent attachment of [ 14C]methyl groups onto amino and imidazole groups by reaction in vacuo with [14C]methyl iodide. The method was tested by investigating the biodistribution of 14C in rats that were fed 14C-labeled human soluble cluster of differentiation 14 (CD14) protein, a receptor for bacterial lipopolysaccharide. Two other control proteins, bovine serum albumin (BSA) and casein, were also labeled with 14C and used for comparative analysis to determine the following: (i) the efficacy and cost efficiency of the in vacuo radiolabeling procedure and (ii) the extent of incorporation of the 14C label into the organs of orogastrically fed 10-day-old Sprague-Dawley rats. [14C]BSA, [ 14C]casein, and [14C]CD14 were individually prepared with specific radioactivities of 34,400, 18,800, and 163,000 disintegrations per minute (dpm)/μg, respectively. It was found that the accumulation of 14C label in the organs of [14C]CD14-fed rats, most notably the persistence of 14C in the stomach 480 min postgavage, was temporally and spatially distinct from [14C]BSA and [ 14C]casein-fed rats. © 2010 Elsevier Inc. All rights reserved.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Steacie Institute for Molecular Sciences (SIMS-ISSM)
Peer reviewedYes
NPARC number21271362
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Record identifierf98e8f9a-e3f5-4081-9ef6-6c239672a029
Record created2014-03-24
Record modified2016-05-09
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