A chip-based miniaturized format for protein-expression profiling: the exploitation of comprehensively produced antibodies.
Numerous antibodies have been developed and validated in recent years, and show promise for use in novel functional protein assays. Such assays would be an alternative to pre-existing comprehensive assays, such as DNA microarrays. Antibody microarrays are thought to represent those functional protein assays. While a variety of attempts have been made to apply DNA microarray technology to antibody microarrays, a fully optimized protocol has not been established. We have been conducting a project to comprehensively produce antibodies against mouse KIAA ("KI" stands for "Kazusa DNA Research Institute" and "AA" are reference characters) proteins. Using our library of antibodies, we established a novel antibody microarray format that utilizes surface plasmon resonance (SPR) technology. A label-free real-time measurement of protein expression in crude cell lysates was achieved by direct readout of the bindings using SPR. Further refinement of the antibody microarray format enabled us to detect a smaller quantity of target proteins in the lysate without the bulk effect. In this review, we first summarize available antibody array formats and then describe the above-mentioned format utilizing updated SPR technology.
Koga
Hisashi
H
Chiba Industry Advancement Center, Chiba, Japan. hkoga@kazusa.or.jp
Kyo
Motoki
M
Usui-Aoki
Kazue
K
Inamori
Kazuki
K
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008904:Miniaturization; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity
D000906:Antibodies; D011506:Proteins