Characterization of monoclonal antibodies specific to human tartrate-resistant acid phosphatase.
A major product of osteoclasts, tartrate-resistant acid phosphatase (TRAP) is an essential but insufficient enzyme for bone resorption. TRAP is an excellent cell marker for osteoclasts and macrophages and is being investigated as a serum marker for osteoclast activity in diseases of bone destruction. For decades, TRAP has also been used as a marker for hairy cell leukemia. Immunoassays for TRAP are sought to increase the sensitivity and specificity of the TRAP test for bone and hairy cells. Our laboratory recently developed a monoclonal antibody to TRAP (9C5) useful for immunohistochemical identification of TRAP-positive cells in paraffin sections. Herein, we characterize 9C5 in greater detail and report production of another anti-TRAP monoclonal antibody antibody (14G6) reactive with native, active enzyme antigen. Enzyme immunoassay, immunoprecipitation, western blot, and immunohistochemical analyses revealed the contrasting properties of 9C5 and 14G6. Antibody 9C5 reacts with a heat-denatured epitope and is suitable for denaturing western blot analysis and for immunohistochemistry. Antibody 14G6 reacts with a conformational determinant destroyed by heat and is suitable for immunoprecipitation of active TRAP, although 20% to 30% of activity is inhibited in the immune complexes. Having characterized several properties of these anti-TRAP antibodies, 9C5 and 14G6 may be useful for development of TRAP-specific immunoassays in bone pathology and hematology. They will certainly be of use for the study of biosynthesis, regulation, expression, and function of TRAP.
Janckila
A J
AJ
Special Hematology Laboratory, Veterans Affairs Medical Center, Louisville, Kentucky, USA.
Cardwell
E M
EM
Yam
L T
LT
D000135:Acid Phosphatase / Q000276:immunology*; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015415:Biomarkers; D015153:Blotting, Western; D001862:Bone Resorption / Q000201:enzymology / Q000276:immunology; D000939:Epitopes; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods*; D007527:Isoenzymes / Q000276:immunology*; D007943:Leukemia, Hairy Cell / Q000201:enzymology / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D009000:Monocytes / Q000276:immunology; D000071681:Tartrate-Resistant Acid Phosphatase
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D000939:Epitopes; D007527:Isoenzymes; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase