A novel approach to protein expression profiling using antibody microarrays combined with surface plasmon resonance technology.
We have previously described our systems for the high-throughput production of antibodies against mouse KIAA proteins and their validation (Proteomics 2004, 4, 1412-1416). Using our "libraries" of antibodies, we established a novel antibody microarray system in which surface plasmon resonance (SPR) technology is utilized for signal detection. Up to 400 real-time antibody-target bindings could be measured simultaneously within a single hour. This rapid detection was achieved by direct readout of the bindings using SPR technology. To evaluate our system, we assessed the reproducibility on crude protein samples and obtained satisfactorily reproducible results, exhibiting correlation values >0.92. Using this SPR-based antibody microarray system, we examined mKIAA protein expression in five different adult mouse tissues and identified the specific tissue expression patterns of several mKIAA proteins.
Usui-Aoki
Kazue
K
Chiba Industry Advancement Center, Nakase, Mihama-ku, Chiba, Japan.
Shimada
Kiyo
K
Nagano
Mihoko
M
Kawai
Makoto
M
Koga
Hisashi
H
D000818:Animals; D000906:Antibodies; D051379:Mice; D040081:Protein Array Analysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics; D011817:Rabbits; D011994:Recombinant Proteins / Q000032:analysis; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance; D014018:Tissue Distribution
D000906:Antibodies; D011506:Proteins; D011994:Recombinant Proteins