Endogenous allergen assessments - Europabio

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Considerations for Serology Based Studies 6 Primary Limitation – limited availability of well-characterized sera. - Variability in IgE binding across one or more proteins among different patients. • Not all patients have IgE against a given <strong>allergen</strong> of the same affinity – thus, quantitative determinations are limited. - Soybean has many <strong>allergen</strong>s, so thoroughly characterizing the exact same set of proteins with good reproducibility from one study to the next is not possible since sera will vary temporally among patients. - Virtually impossible to have same set of patient sera form one study to next
Liquid Chromatography MS/MS for Protein Identification 7 1. Proteins are digested into peptides 2. Peptides are separated to reduce complexity 3. Peptides are “weighed” and sequenced by mass spectrometry 4. Peptide sequence provides protein identification through bioinformatics S.E. Stevenson, Houston N., and Thelen J. 2010. Regul. Toxicol. Pharmacol. 58, 3, S36-41.
Page 1 and 2: Design, relevance and limitations o
Page 3 and 4: IgE, ug/ml Question: Are Endogenous
Page 5: 2-D Electrophoresis and Western Blo
Page 9 and 10: Aqua® Peptides and Quantifying; Ho
Page 11 and 12: µg allergen protein/mg total prote
Page 13 and 14: Soybeans 13 Glycine max Yes, it’s
Page 15 and 16: Relevance of the data - Can this da
Page 17: Questions and Discussion ● Thank

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