Isolation of Patient-Related Antigens from Allergen Extracts by Means of Protein A-Sepharose-Patient IgG1,2,4 Bio-Affinity Chromatography

Authors

  • I. Søndergaard,

    Corresponding author
    1. Medical Department TA, Division of Allergology and Laboratory for Medical Allergology, Rigshospital, University of Copenhagen, Copenhagen, Denmark
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  • B. Weeke

    1. Medical Department TA, Division of Allergology and Laboratory for Medical Allergology, Rigshospital, University of Copenhagen, Copenhagen, Denmark
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Address: Ib Søndergaard, Laboratory for Medical Allergology 7542, Rigshospitalet, Tagensvej 20, DK-2200 Copenhagen N, Denmark

Abstract

A method is described for bio-affinity purification of patient-specific IgG reactive antigens by using protein A-sepharose allergic patient serum and the corresponding antigen/allergen extract. The optimal conditions for each step have been determined and it is shown that the non-specific binding of both serum proteins and antigens can be kept at very low levels by a high ionic strength. The specificity of the reaction has been shown by biospecific elution of 125I-labelled antigens by non-labelled antigens and also by comparison of the amount of antigen bound by sera from hyposensitized patients with that from non-atopic individuals. It is suggested that the isolated antigen-IgG-immune complexes could be used for a specific and active immunotherapy.

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