Quantitative immunohistochemistry using an antibody-fused bioluminescent protein
Ke-Yong Wang,
Chun Wu,
Shohei Shimajiri,
Toshiteru Enomoto,
Hidehiro Kubota,
Hidefumi Akiyama,
Yoshihiro Ohmiya
Affiliations
Ke-Yong Wang
1Shared-Use Research Center, School of Medicine, University of Occupational & Environmental Health, Iseiga-oka 1-1, Kita-kyusyu 807-8555, Japan
Chun Wu
2Biomedical Research Institute, National Institute of Advanced Industrial Science & Technology (AIST), Midorioka 1-8-31, Ikeda, Osaka 563-8577, Japan
Shohei Shimajiri
3Department of Surgical Pathology, School of Medicine, University of Occupational & Environmental Health, Iseiga-oka 1-1, Kita-kyusyu 807-855, Japan
Toshiteru Enomoto
4ATTO Corporation, 3-2-2 Motoasakusa, Taito-ku, Tokyo 111-0041
Hidehiro Kubota
4ATTO Corporation, 3-2-2 Motoasakusa, Taito-ku, Tokyo 111-0041
Hidefumi Akiyama
5Tokyo University Institute for Solid State Physics, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8581, Japan
Yoshihiro Ohmiya
7DAILAB, DBT-AIST International Center for Translational & Environmental Research (DAICENTER), National Institute of AIST, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan
We established a quantitative detection method for immunohistochemistry based on a reference standard light-emitting diode, protein microarray and antibody-fused bioluminescent protein. In this procedure, we calibrated the bioluminescence imaging system and prepared the calibration curve between antigen and antibody-fused bioluminescent protein using a protein microarray. Then we converted the detecting light signal to antigen count via absolute photon number in the bioluminescent images; there was a resulting threefold difference in the target antigen number between normal and cancerous tissues. Our technique can easily compare immunohistological images and evaluate tumor progression in quantitative pathological diagnosis.
