The metabolic reactions by cytochrome P450 (P450) are critical to evaluate toxicity of xenobiotics. We developed a biochip that can assay multiple P450 species in parallel by immobilizing P450s in microwells (diameter and depth: 50�m) fabricated on a cyclo-olefine polymer (COP) plate. P450s were printed into microwells by the inkjet printing together with agarose and subsequently immobilized by the gelation of agarose. To minimize the activity loss due to rapid dehydration, we added trehalose or albumin (bovine serum) to the P450 solution. Assays were done by incorporating substrate solutions into microwells. We regulated the initiation of P450 reactions by using photo-protected (caged) glucose-6-phosphate, which is required for generating the cofactor of P450 (NADPH). It enabled a more controlled and quantitative assay of P450 reactions in microwells. By combining microwell configuration, inkjet printing technology, and caged compounds, we could establish a platform for high-throughput assay of P450s.

P450 microarrays fabricated in microwells by the inkjet printing

243rd National Meeting (2012)

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