Generation and characterization of monoclonal antibodies to human type-5 tartrate-resistant acid phosphatase: development of a specific immunoassay of the isoenzyme in serum.
We have characterized four monoclonal antibodies (mAbs) to the purple ("tartrate-resistant," band 5) acid phosphatase of the human osteoclast (TRAP) and used these to develop a specific serum immunoassay. All four mAbs are of high affinity (Ka = 1-5 x 10(8) L/mol) with a very fast Kassoc (0.2-2.0 x 10(5) L mol-1 s-1) and a moderate Kdissoc (1-3 x 10(-3) s). Two of the mAbs were selected to develop a time-resolved fluorescence immunoassay to measure serum concentrations of TRAP. The mean serum immunoreactive TRAP in a group of healthy premenopausal women and men was 3.7 +/- 1.8 micrograms/L (mean +/- SD) and 3.5 +/- 1.6 micrograms/L, respectively. Significantly higher concentrations of TRAP were found in postmenopausal women (6.3 +/- 2.3 micrograms/L) and in eight patients with Gaucher disease (19.3 +/- 4.7 micrograms/L). Further studies are required to investigate the value of serum TRAP as a marker of bone resorption.
Chamberlain
P
P
SmithKline Beecham Pharmaceuticals, Epsom, Surrey, UK.
Compston
J
J
Cox
T M
TM
Hayman
A R
AR
Imrie
R C
RC
Reynolds
K
K
Holmes
S D
SD
D000135:Acid Phosphatase / Q000097:blood* / Q000276:immunology*; D000293:Adolescent; D000328:Adult; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000915:Antibody Affinity; D015415:Biomarkers; D001847:Bone Diseases / Q000201:enzymology; D001862:Bone Resorption; D005260:Female; D015200:Fluoroimmunoassay / Q000379:methods* / Q000706:statistics & numerical data; D005776:Gaucher Disease / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000097:blood* / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D017698:Postmenopause; D017697:Premenopause; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D000071681:Tartrate-Resistant Acid Phosphatase
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D007527:Isoenzymes; D011994:Recombinant Proteins; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase