Establishing objective detection limits for the pepsin digestion assay used in the assessment of genetically modified foods

A. O. Ofori-Anti, H. Ariyarathna, L. Chen, H. L. Lee, S. N. Pramod, R. E. Goodman

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Rationale: Guidelines for assessing the potential allergenicity of genetically modified (GM) organisms recommend testing the digestibility of the introduced protein by pepsin. Previous studies detailed the digestion procedure but have not described a simple objective measurement of the extent of digestion nor evaluated the impact of variation in pepsin activity. Methods: Samples of eight proteins were digested by pepsin at pH 1.2 and 2.0 using standard conditions (10,000 U of pepsin activity per mg test protein) as well as 5000 and 20,000 units per mg of test protein. An independent digestion assay of hemoglobin was used to verify pepsin activity for each assay. Digestion was stopped in timed samples between 0.5 and 60 min. Digestion samples and undigested protein (10% and 100%) were separated by SDS-PAGE. Residual stained protein bands were measured by image analysis. Results: The differences in pH and pepsin concentration only had minor effects on digestion of intermediately stable proteins: concanavalin A, ovalbumin, and lysozyme, but not on rapidly digested or stable proteins. Conclusions: Verification of pepsin activity and measurement of an objective endpoint of digestion (e.g. (90%) should provide more comparable results for the safety assessment of novel food proteins.

Original languageEnglish (US)
Pages (from-to)94-103
Number of pages10
JournalRegulatory Toxicology and Pharmacology
Volume52
Issue number2
DOIs
StatePublished - Nov 1 2008

Fingerprint

Genetically Modified Food
Pepsin A
Limit of Detection
Digestion
Assays
Proteins
Genetically Modified Organisms
Ovalbumin
Muramidase
Concanavalin A
Image analysis
Polyacrylamide Gel Electrophoresis
Hemoglobins
Guidelines

Keywords

  • Allergenicity
  • Food allergy
  • Genetically modified
  • Pepsin digestion
  • Simulated gastric fluid

ASJC Scopus subject areas

  • Toxicology

Cite this

Establishing objective detection limits for the pepsin digestion assay used in the assessment of genetically modified foods. / Ofori-Anti, A. O.; Ariyarathna, H.; Chen, L.; Lee, H. L.; Pramod, S. N.; Goodman, R. E.

In: Regulatory Toxicology and Pharmacology, Vol. 52, No. 2, 01.11.2008, p. 94-103.

Research output: Contribution to journalArticle

Ofori-Anti, A. O. ; Ariyarathna, H. ; Chen, L. ; Lee, H. L. ; Pramod, S. N. ; Goodman, R. E. / Establishing objective detection limits for the pepsin digestion assay used in the assessment of genetically modified foods. In: Regulatory Toxicology and Pharmacology. 2008 ; Vol. 52, No. 2. pp. 94-103.
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