51. Quantitative immunohistochemistry using an antibody-fused bioluminescent protein
- Author
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Hidefumi Akiyama, Chun Wu, Toshiteru Enomoto, Yoshihiro Ohmiya, Ke-Yong Wang, Shohei Shimajiri, and Hidehiro Kubota
- Subjects
0303 health sciences ,biology ,040301 veterinary sciences ,Chemistry ,Protein Array Analysis ,04 agricultural and veterinary sciences ,Molecular biology ,Immunohistochemistry ,General Biochemistry, Genetics and Molecular Biology ,Antibodies ,0403 veterinary science ,03 medical and health sciences ,Luminescent Proteins ,Antigen ,Tumor progression ,biology.protein ,Protein microarray ,Bioluminescence imaging ,Bioluminescence ,Humans ,Luciferase ,Antibody ,030304 developmental biology ,Biotechnology - Abstract
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.
- Published
- 2020