Celiac antigen
AssigneeXybernaut Corporation
InventorAmin Tjota
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Abstract
In this invention a novel antigen is provided for screening patients suspected of having celiac disease. The antigen is prepared by starting with human placenta tissue which is perfused with a herpes and collagenase buffer to obtain a signle cell suspension. This suspension is enriched to obtain an enriched protein portion and then separated out to obtain an embryonic celiac antigen (ECA). This ECA is used with serum from patients to effect binding of ECA with IgA in the serum while applying a human IgA antibody to the serum. The results are then read on a spectrophotometer to confirm or negate the presence of celiac disease.
Claims1
A method for the isolation of an antigen from human placenta tissue to be used as a screening assay for celiac disease comprising:
- a) cleaning and washing said placenta tissue with HEPES buffer;
- b) digesting the said placenta tissue with collagenase buffer to obtain a placenta cell population in suspension, said placenta cell population containing cytotrophoblast cells;
- c) enriching the cytotrophoblast cell population in said suspension via a 0-30% discontinuous Percoll gradient;
- d) extracting protein from said enriched cytotrophoblast cells with lysing buffer, extraction buffer and via ultracentrifugation to obtain a protein portion;
- e) isolating embryonic celiac antigen (ECA) from said extracted protein by applying said protein portion to an ion exchange DEAE Sephadex column followed by size exclusion gel chromatography;
- and
- f) identifying by SDS-PAGE and western blotting embryonic celiac antigens comprising protein molecules with an isoelectric range (pI) of from 5.1-5.8 and an apparent molecular weight selected from the group consisting of 55 kd, 65 kd, and 110 kd.
Dependencies are inferred from the claim text.