Characterization of acute renal allograft rejection by human serum proteomic analysis.
To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS) were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups: acute rejection (AR), stable renal function (SRF) and normal volunteer (N). Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.
Gao
Ying
Y
Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Key Laboratory of Organ Transplantation, Ministry of Education, Ministry of Health, Wuhan, 430030, China. gaotongji@163.com
Wu
Ke
K
Xu
Yi
Y
Zhou
Hongmin
H
He
Wentao
W
Zhang
Weina
W
Cai
Lanjun
L
Lin
Xingguang
X
Fang
Zemin
Z
Luo
Zhenlong
Z
Guo
Hui
H
Chen
Zhonghua
Z
D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000097:blood; D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D006084:Graft Rejection / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D016030:Kidney Transplantation / Q000009:adverse effects*; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D044967:Serum / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D014809:Vitamin D-Binding Protein / Q000097:blood; D055815:Young Adult
D015415:Biomarkers; D020543:Proteome; D014809:Vitamin D-Binding Protein