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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Published on: July 31, 2011
Immunochemical and biological quantification of peanut extract
Lars K Poulsen1, Mona H Pedersen, Michael Platzer
1Lab. Medical Allergology 7542, National University Hospital, Blegdamsvej 9, DK-2100 Copenhagen.
Arbeiten Aus Dem Paul-Ehrlich-Institut (Bundesamt Fur Sera Und Impfstoffe) Zu Frankfurt A.M
|May 4, 2004
Summary
Rabbit basophils offer a viable ex vivo method for biological standardization of allergen extracts. This histamine release assay provides a valuable tool for characterizing allergen potency, complementing existing methods.
Area of Science:
- Immunology
- Allergology
- Biotechnology
Background:
- Biological standardization of allergen extracts is crucial for characterizing their potency.
- Skin prick testing is the gold standard, but histamine release (HR) offers a convenient ex vivo alternative for high-throughput analysis.
Purpose of the Study:
- To evaluate the utility of rabbit basophils for biological standardization of allergen extracts.
- To compare the sensitivity of rabbit basophil histamine release with other methods, including ELISA and human basophil assays.
Main Methods:
- Peanut allergen was used as a model.
- Rabbit basophils were sensitized directly via immunization or passively using rabbit antiserum.
- Histamine release experiments were conducted on sensitized rabbit and human basophils.
- Results were compared to an ELISA assay.
Main Results:
- Directly sensitized rabbit basophils showed histamine release upon allergen exposure.
- Passively sensitized rabbit basophils were less sensitive (100-1000 times lower) than directly sensitized cells.
- Overall sensitivity ranking: ELISA > HR-human cells > HR-sensitized rabbit cells > HR-passively sensitized rabbit cells.
Conclusions:
- Rabbit basophils can be effectively utilized for biological standardization of allergen extracts.
- This method enables the use of rabbit antisera in standardization processes.
- The sensitivity of rabbit basophil assays is lower than ELISA and human basophil assays but offers a valuable alternative.
