Determining allergen sensitization in models

If you are using an allergen, such as house dust mite (HDM) material, in your animal model and you want to analyse the stage of sensitization we can offer you the information and tools to do so.

The serum of a model will naturally contain a myriad of different proteins and antibodies . In order to investigate the stage to which the model has been sensitized or, in a later phase, how the model responds to treatment you need to quantify the (allergen specific) antibodies. This requires an immunoassay that is of highly specific that can be used on a complex sample like serum.

Assay Materials

A method that suits these criteria is a sandwich enzyme-linked immunosorbent assay (ELISA). With this method an ELISA plate is coated with a capture antibody which for this purpose are anti-(model animal) antibodies. For example anti-mouse IgE, IgG1 or IgG2a. Once the sample is added the antibodies in the serum will bind to the anti-antibodies of choice on the plate, the rest of the serum is subsequently washed away due to being unbound. In a regular sandwich ELISA a detection antibody would now be added to quantify the amount of bound substrate however Citeq Biologics has biotinylated products on offer that can act as a more clinically relevant capture antibody. This saves having to purify the specific antibodies and then having to detect if they have the specific affinity for the allergen as well.

• Biotinylated D. pteronyssinus (HDM) extract
• Biotinylated Der p 1
• Biotinylated Der p 2
• Biotinylated D. farinae (HDM) extract

More detailed information regarding biotinylation can be found here. With these biotinylated products it becomes possible to quantify the IgE, IgG1 or IgG2a response to house dust mite or their major allergens specifically. This reduces some of the possible variables when quantifying the immune response and by allowing you to measure the response to treatment in greater detail it may be possible to uncover new insights that previously might have been overlooked.

A good example of the use of this type of measuring can be found in a paper by Leonie S. van Rijt. They used a mouse model centred around  birch pollen immunotherapy and the inhibition of Th2 inflammation. With the ability to look at allergen-specific immunoglobins they not only were able to determine beyond a doubt their mice were correctly sensitized, by quantifying allergen-specific immunoglobins they were able to determine  that the gradual reduction in AHR in their model was inversely associated with the increase of IgG2a.

The anti-mouse IgE, IgG1 or IgG2a are a great tool to quantify the immune activity in your model and in combination with biotinylated products it becomes possible to gather even more information.