Development of immunoassays for serum tartrate-resistant acid phosphatase isoform 5a

Serum tartrate-resistant acid phosphatase (TRACP) consists of 2 structurally related isoforms, TRACP 5a and 5b. TRACP 5b is from bone-resorbing osteoclasts. TRACP 5a may be a macrophage product of inflammation. We used a novel antibody to TRACP 5a to standardize immunoassays for serum TRACP 5a activ...

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Bibliographic Details
Published in:Clinica chimica acta 2005-09, Vol.359 (1), p.132-140
Main Authors: Chao, Tsu-Yi, Lee, Su-Huei, Chen, Mary M., Neustadt, David H., Chaudhry, Uzma A., Yam, Lung T., Janckila, Anthony J.
Format: Article
Language:English
Subjects:
Acid phosphatase
Acid Phosphatase - blood
Acid Phosphatase - immunology
Adult
Aged
Arthritis, Rheumatoid - blood
Bone resorption
Female
Humans
Immunoassay
Immunoassay - methods
Inflammation
Isoenzymes - blood
Isoenzymes - immunology
Male
Middle Aged
Tartrate-Resistant Acid Phosphatase
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Summary:Serum tartrate-resistant acid phosphatase (TRACP) consists of 2 structurally related isoforms, TRACP 5a and 5b. TRACP 5b is from bone-resorbing osteoclasts. TRACP 5a may be a macrophage product of inflammation. We used a novel antibody to TRACP 5a to standardize immunoassays for serum TRACP 5a activity and protein. Biotinylated anti-TRACP antibodies were used to immobilize serum TRACP isoforms. TRACP activity was measured using 4-nitrophenyl phosphate as substrate. TRACP 5a protein was measured with an independent peroxidase-conjugated anti-TRACP antibody. Immunoassays were standardized for linearity of serum dose response, sensitivity and precision. Reference ranges for TRACP 5a were established from serum of 50 healthy males and 50 healthy age-matched females. Serum TRACP 5a activity and protein were determined in 29 cases of rheumatoid arthritis. Serum matrix interference in both TRACP 5a assays required dilution to 10% serum to approach linearity. Intra-assay and inter-assay CV% were
ISSN:0009-8981
1873-3492
DOI:10.1016/j.cccn.2005.03.039