Generation and characterization of putative monoclonal and polyclonal antibodies against tartrate-resistant acid phosphatase.
The generation of monoclonal antibodies (MAbs) specific for tartrate-resistant acid phosphatase (TRAP) may aid development of a serological immunoassay for this marker of bone resorption. The lack of MAbs to TRAP largely reflects the difficulty in obtaining sufficient antigen for in vivo immunization strategies. We have circumvented this problem by using in vitro immunization, requiring a small amount of TRAP isolated from osteoclastoma tumor. Two MAbs designated 312D and 310A were generated that exhibited weak anti-TRAP activity in enzyme-linked immunosorbent assay and immunocytochemistry. A polyclonal antibody to TRAP (Ab8023) was also raised in rabbit, using synthetic peptide. Ab8023 and MAbs 310A and 312D exhibited no activity against TRAP in dot-blotting experiments. Further characterization in enzyme-liked immunosorbent assay showed that Ab8023 was remarkably specific for TRAP whereas MAb 312D cross-reacted with another metalloenzyme, human prostatic acid phosphatase.
Nelson
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PN
Department of Histopathology, St. George's Hospital Medical School, Tooting, London, U.K.
Chambers
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T
D000135:Acid Phosphatase / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000917:Antibody Formation; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000276:immunology*; D008297:Male; D051379:Mice; D011467:Prostate / Q000201:enzymology; D011817:Rabbits; D000071681:Tartrate-Resistant Acid Phosphatase; D013644:Tartrates / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D013644:Tartrates; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase; C029768:tartaric acid