Cross-reactivity studies staining methods

As mentioned earlier, the purpose of this review is not to detail the actual methods used for tissue cross-reactivity studies but rather to highlight the many important factors that must be taken under consideration during the conduct, evaluation, and regulatory review and of the study. Specific details for tissue collection, generation of control materials, and staining procedures themselves are readily available in the literature. 

The following factors should be considered when choosing a staining method for a tissue cross-reactivity study (1) test article forms that are available, (2) test article affinity, (3) proposed plasma level for clinical studies, (4) epitope stability, and (5) ability to scale up the method to stain a large number of tissues. These various issues are addressed below. 

As mentioned earlier, there are other ways to label antibodies for use in tissue cross-reactivity studies. Detailed discussion of various labels and staining methods associated with them are beyond the scope of this review. However,... 

Staining adequacy is addressed by staining replicate cryosections from each tissue for an antigen known to be expressed on components common to all tissues (e.g., CD31, [T-microglobulin, transferrin receptor), as discussed previously. This ensures that the tissue can be stained using immunohistochemical methods and thus is a suitable sample for the tissue cross-reactivity study. 

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