Quantification of Chromogranin A and Its Fragments in Biological Fluids

Human chromogranin A (CgA), a 439-residue long neurosecretory protein, can serve as a circulating biomarker for a wide range of neuroendocrine tumors. Increased levels of immunoreactive CgA are also present in the blood of patients with cardiovascular, gastrointestinal, or inflammatory diseases with, in certain cases, important diagnostic and prognostic implications. A growing body of evidence suggest that CgA and various CgA-derived fragments have complex roles in the regulation of cardiovascular system, metabolism, innate immunity, angiogenesis, and tissue repair, sometime with opposite biological effects. For example, while full-length CgA (CgA_1-439) inhibits angiogenesis, the CgA_1-373 fragment, at certain doses, is proangiogenic. Thus, the selective quantification of CgA and its fragments in the blood of patients (and in other biological fluids) is of great experimental and clinical interest. Here, we describe methods to produce CgA_1-439 and CgA_1-373 and to develop ELISAs capable of detecting these polypeptides in a very selective manner. The same approach can be used, in principle, also for developing assays for other fragments.