IMMUNOASSAYS AND METHODS OF DETECTING AND MEASURING INTACT FIBROBLAST GROWTH FACTOR 23, AND C-TERMINAL AND N-TERMINAL FRAGMENTS THEREOF

摘要

Immunoassays and methods for detecting and quantifying biological levels of intact fibroblast growth factor (FGF)-23, as well as the N-terminal and C-terminal fragments thereof in a biological sample. The relative amounts or ratios of FGF-23 relative the N-terminal and C-terminal fragments can also be determined. The systems and methods deploy antibodies that are specific to antigenic regions formed upon either the N-terminal or C-terminal regions of FGF-23 and are systematically applied such that intact FGF-23 and the fragments thereof can be detected and quantified. In certain embodiments, dissimilar labels conjugated to tracer antibodies or labeled antibodies specific to N-terminal and/or C-terminal tracer antibodies are utilized to facilitate detection and quantification of both whole length FGF-23 and any fragments thereof.

主权项

1. An immunoassay for detecting intact FGF-23 protein molecules and C-terminal fragments thereof, and further identifying the relative amounts of said intact FGF-23 and said C-terminal fragments, said immunoassay comprising:
a C-terminal capture antibody, said C-terminal capture antibody having an affinity for and specifically binding to an antigenic region falling within amino acids 180-251 of FGF-23 (SEQ ID No.: 4); a C-terminal tracer antibody having a label conjugated therewith, said C-terminal tracer antibody having an affinity for and specifically binding to an antigenic sequence falling within amino acids 180-251 of FGF-23 (SEQ ID NO.: 4), said C-terminal tracer antibody having an affinity for and specifically binding to an antigenic sequence dissimilar to that of said C-terminal capture antibody such that in use, said C-terminal capture antibody and said C-terminal tracer antibody are capable of sequentially and simultaneously binding to the respective antigenic regions to which each respective antibody specifically binds; an N-terminal tracer antibody having a label conjugated therewith, said N-terminal tracer antibody having an affinity for and specifically binding to an antigenic region formed within amino acids 25-179 of FGF-23 (SEQ ID NO.: 3), said N-terminal tracer antibody being able to bind to said antigenic region sequentially or simultaneously in combination with said C-terminal tracer antibody and said C-terminal capture antibody.