Development of retinol-binding protein 4 immunocolloidal gold fast test strip using high-sensitivity monoclonal antibodies generated by DNA immunization.
DNA immunization is an efficient method for high-affinity monoclonal antibody generation. Here, we describe the generation of several high-quality monoclonal antibodies (mAbs) against retinol-binding protein 4 (RBP4), an important marker for kidney abnormality and dysfunction, with a combination method of DNA priming and protein boost. The mAbs generated could bind to RBP4 with high sensitivity and using these mAbs, an immunocolloidal gold fast test strip was constructed. The strip can give a result in <5 min and is very sensitive with a detection limit of about 1 ng/ml. A small-scale clinical test revealed that the result of this strip was well in accordance with that of an enzyme-labeled immunosorbent assay kit currently available on the market. Consequently, it could be useful for more convenient and faster RBP4 determination in the clinic.
Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, China. cbian@sibs.ac.cn
Chao
C
Bian
Fang
F
Zhang
Feng
F
Wang
Zhiyang
Z
Ling
Min
M
Luo
Hongqiang
H
Wu
Yizhuo
Y
Sun
Junhui
J
Li
Bingnan
B
Li
Jingyan
J
Zhu
Linna
L
Tang
Yanyan
Y
Zhou
Qunfang
Q
Shi
Yongyong
Y
Ji
Lin
L
Tian
Guomei
G
Lin
Yu
Y
Fan
Niansong
N
Wang
Bing
B
Sun
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D017979:Gold Colloid; D006801:Humans; D007114:Immunization / Q000379:methods*; D007674:Kidney Diseases / Q000235:genetics / Q000473:pathology*; D011933:Reagent Kits, Diagnostic; D011934:Reagent Strips; D054839:Retinol-Binding Proteins, Plasma / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D017979:Gold Colloid; C101326:RBP4 protein, human; D011933:Reagent Kits, Diagnostic; D011934:Reagent Strips; D054839:Retinol-Binding Proteins, Plasma; D004247:DNA