Monitoring the VEGF level in aqueous humor of patients with ophthalmologically relevant diseases via ultrahigh sensitive paper-based ELISA.
The vascular endothelial growth factor (VEGF) level in aqueous humor has been used as an indicator to monitor specific diseases in the retinal ischemic condition. For clinical diagnosis, only about 200 μL of aqueous humor can be collected from the anterior chamber before the threat of anterior chamber collapse. It is necessary to develop an inexpensive diagnostic approach with the characteristics of highly sensitive, short operation duration, and requires small clinical sample quantities. To achieve the main objective of this study, we first prepared bevacizumab to be conjugated with HRP. We then deposited 2 μL aqueous humor from patients with different diseases onto each test zone of paper-based 96-well plates. After the colorimetric results were performed via ELISA protocol, the output signals were recorded using a commercial desktop scanner for analysis. In this study, only 2 μL from the aqueous humor of each patient was required for paper-based ELISA. The mean aqueous VEGF level was 14.4 pg/mL from thirteen patients (N = 13) with senile cataract as the control. However, the mean aqueous VEGF level from other patients with proliferative diabetic retinopathy (N = 14), age-related macular degeneration (N = 17), and retinal vein occlusion (N = 10) showed VEGF increases to 740.1 pg/mL, 383 pg/mL, and 219.4 pg/mL, respectively.
Department of Ophthalmology, Taichung General Veteran Hospital, Taichung 40705, Taiwan; Institute of Nanoengineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan.
Min-Yen
MY
Hsu
Institute of Nanoengineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan.
Chung-Yao
CY
Yang
Institute of Nanoscience, National Chung Hsing University, Taichung 40227, Taiwan.
Wen-Hsin
WH
Hsu
Department of Ophthalmology, Taichung General Veteran Hospital, Taichung 40705, Taiwan.
Keng-Hung
KH
Lin
Department of Ophthalmology, Taichung General Veteran Hospital, Taichung 40705, Taiwan.
Chun-Yuan
CY
Wang
Department of Ophthalmology, Taichung General Veteran Hospital, Taichung 40705, Taiwan.
Ying-Cheng
YC
Shen
Department of Ophthalmology, Taichung General Veteran Hospital, Taichung 40705, Taiwan.
Yu-Chen
YC
Chen
Department of Ophthalmology, Taichung Tzu Chi Hospital, Taichung 42743, Taiwan.
Siu-Fung
SF
Chau
Department of Ophthalmology, Taichung Tzu Chi Hospital, Taichung 42743, Taiwan.
Hin-Yeung
HY
Tsai
Institute of Nanoengineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 11529, Taiwan. Electronic address: chaomin@mx.nthu.edu.tw.
Chao-Min
CM
Cheng
D001082:Aqueous Humor / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005128:Eye Diseases / Q000378:metabolism*; D006801:Humans; D057230:Limit of Detection; D010209:Paper; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D042461:Vascular Endothelial Growth Factor A
Diabetic disease; Diagnosis; Ophthalmology; Paper-based ELISA; VEGF