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J Immunoassay Immunochem. 2016;37(6):611-22. doi: 10.1080/15321819.2016.1182551.

Development and validation of an enzyme-linked immunosorbent assay for the quantification of gelonin in mouse plasma.

Author information

1
a Department of Pharmaceutical Sciences , School of Pharmacy and Pharmaceutical Sciences, University at Buffalo - The State University of New York , Buffalo , New York , USA.

Abstract

This article details the development and validation of an enzyme-linked immunosorbent assay (ELISA) for the quantification of gelonin in mouse plasma. The ELISA was validated for intra- and inter-day variability and for accuracy over a standard curve range of 7.5-100 ng/mL. The assay was then applied to assess gelonin pharmacokinetics in mice. Results from the ELISA were compared to data obtained from a parallel study conducted with (125)Iodine-labeled gelonin, with quantification via gamma counting. The ELISA demonstrated good precision, as the percent coefficient of variation of quality control samples in intra-day and inter-day validation ranged from 5.4-9.3% and 2.9-7.3%, respectively. Sample recoveries ranged from 98.3-105% of nominal values. The ELISA method yielded lower plasma concentrations of gelonin than found from the less-specific gamma counting method. Consequently, pharmacokinetic analyses yielded significantly higher estimates for volume of distribution (106 ± 31 vs. 55.8 ± 13 mL/kg) and plasma clearance (34.7 ± 6.6 vs. 10.9 ± 2.1 mL/min/kg) for data determined by ELISA vs. by gamma counting.

KEYWORDS:

ELISA; assay; gelonin; pharmacokinetics; protein toxin; ribosome-inactivating protein

PMID:
27135787
DOI:
10.1080/15321819.2016.1182551
[Indexed for MEDLINE]

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