Antibodies to different peptides in osteopontin reveal complexities in the various secreted forms.
We have generated synthetic peptides corresponding to various portions of human osteopontin (OPN) and have immunized rabbits and mice with these peptides to generate polyclonal and monoclonal antibodies specific to human OPN. We then generated six distinct sandwich enzyme-linked immunoabsorbent assay (ELISA) systems by using different pairs of polyclonal and monoclonal antibodies against human OPN. These systems allowed us to detect not only various isoforms and truncated forms of recombinant OPN, but also the glycosylated form of native urinary OPN. Most importantly, tumor-derived OPN was differentially detected by the six ELISA systems. The ELISA systems that we have developed will be useful for clarifying the functional roles for OPN in vivo in various physiologic and pathologic conditions.
Kon
S
S
Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Japan.
Maeda
M
M
Segawa
T
T
Hagiwara
Y
Y
Horikoshi
Y
Y
Chikuma
S
S
Tanaka
K
K
Rashid
M M
MM
Inobe
M
M
Chambers
A F
AF
Uede
T
T
D000328:Adult; D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D015153:Blotting, Western; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008297:Male; D051379:Mice; D053495:Osteopontin; D010957:Plasmids; D020033:Protein Isoforms; D012313:RNA / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000737:chemistry* / Q000276:immunology* / Q000652:urine; D013917:Thrombin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D020033:Protein Isoforms; D011994:Recombinant Proteins; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D053495:Osteopontin; D012313:RNA; D013917:Thrombin