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Methods and compositions for protein-hydroxy apatite complexes and their application in testing and modulating immunological system including a novel in vitro test for the detection of antibodies against calcium binding protein-hydroxy apatite complexes
Methods and compositions for protein-hydroxy apatite complexes and their application in testing and modulating immunological system including a novel in vitro test for the detection of antibodies against calcium binding protein-hydroxy apatite complexes
The invention relates to methods and compositions for the manufacture and use of novel hydroxy apatite (HA)-calcium binding protein (CaBP) complexes (CaBP-HA complex) that are useful as antigen preparations for immunological assays for the diagnosis, prognosis, and monitoring of mammalian diseases. The CaBP-HA complexes may be manufactured using synthetic HA that is subjected to serum proteins or by harvesting nanobacteria (NB), also called calcifying nano-particles (CNP), from a mammal. The hydroxy apatite is prepared by incubation with appropriate proteins resulting in conformational changes in said bound proteins. Secondary conformational changes occur when said CaBP-HA complex is subjected to enzymes such as transglutaminase thereby creating covalently bound neoepitopes. The CaBP-HA complex may also contain lipopolysaccharide binding protein (LPSBP) providing for anti-CaBP-LPSBP-HA antibodies detection indicative of disease. Detection of anti-CaBP-HA antibodies in a mammal is via enzyme linked immunosorbent assay.
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