Tunable two dimensional protein patterns through self-assembly nanosphere template.
By the aim of constructing surfaces for multi-component and multifunctional bioassay, a microsphere lithography technique was employed to control the surface morphology. Two kinds of protein molecules (antibodies) were used as building blocks. As a result, dual-component biocompatible surfaces with alternate immunoglobulin micropatterns were fabricated. The employed antibodies included human Immunoglobulin G (IgG) and rabbit IgG, which composed nanometer scale surface arrays on the surfaces. The antibodies were identified specially by immunoreactions with labeled antigens of fluorescein isothiocyanate (FITC)-antihuman IgG and tetramethylrhodamine-5-(and 6)-isothiocyanate (TRITC)-antirabbit IgG. The immune responses were confirmed by confocal fluorescence (FL) microscopy. A study on the sensitivity and quantification was done by using surface-enhanced resonance Raman scattering (SERRS) spectroscopy. The obtained SERRS spectra showed satisfactory resolution in the multi-component detection objects. No interference was observed from inner- or interactions of detecting molecules. The detection limits for both of the antigens reached to as low as 1 ng/mL, which was comparable to FL method. Meanwhile, a good linear relationship between SERRS peak intensity and the logarithm of antigens' concentrations (from 1 ng/mL to 1 mg/mL) were observed. The results demonstrated that SERRS is a very promising detection technique for multi-component immunoassay, and has great potential applications in biotechnology and biochemistry.
State Key Laboratory of Supramolecular Structure and Materials, Jilin University, Changchun 130012, People's Republic of China.
Zhishi
Z
Li
Weidong
W
Ruan
Shanshan
S
Shen
Haiyang
H
Wang
Zhinan
Z
Guo
Xiangxin
X
Xue
Zhu
Z
Mao
Wei
W
Ji
Xu
X
Wang
Wei
W
Song
Bing
B
Zhao
D000818:Animals; D000936:Antigen-Antibody Complex / Q000737:chemistry / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D054118:Nanospheres / Q000737:chemistry* / Q000648:ultrastructure; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D011817:Rabbits; D012235:Rhodamines / Q000737:chemistry / Q000378:metabolism; D012996:Solutions; D013059:Spectrum Analysis, Raman
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D011506:Proteins; D012235:Rhodamines; D012996:Solutions; C009434:tetramethylrhodamine isothiocyanate; D016650:Fluorescein-5-isothiocyanate