An auto-biotinylated bifunctional protein nanowire for ultra-sensitive molecular biosensing.
In order to obtain an ultra-sensitive molecular biosensor, we designed an auto-biotinylated bifunctional protein nanowire (bFPNw) based on the self-assembly of a yeast amyloid protein, Sup35, to which protein G and a biotin acceptor peptide (BAP) were genetically fused. These auto-biotinylated bFPNws can transfer hundreds of commercially available diagnostic enzymes to an antigen-antibody complex via the biotin-avidin system, greatly enhancing the sensitivity of immune-biosensing. Compared to our previously reported seeding-induced bFPNws (Men et al., 2009), these auto-biotinylated bFPNws gave greater signal amplification, reduced non-specific binding and improved stability. The auto-biotinylated self-assembled bFPNw molecular biosensors were applied to detect Yersinia pestis (Y. pestis) F1 antigen and showed a 2000- to 4000-fold increase in sensitivity compared to traditional immunoassays, demonstrating the potential use of these self-assembling protein nanowires in biosensing.
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
Dong
D
Men
Zhi-Ping
ZP
Zhang
Yong-Chao
YC
Guo
Duan-Hao
DH
Zhu
Li-Jun
LJ
Bi
Jiao-Yu
JY
Deng
Zong-Qiang
ZQ
Cui
Hong-Ping
HP
Wei
Xian-En
XE
Zhang
D000942:Antigens, Bacterial / Q000032:analysis; D001363:Awards and Prizes; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods / Q000706:statistics & numerical data; D019921:Biotinylation; D003432:Cross-Linking Reagents / Q000737:chemistry; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods / Q000706:statistics & numerical data; D053770:Nanowires / Q000737:chemistry* / Q000648:ultrastructure; D010454:Peptide Termination Factors / Q000737:chemistry / Q000648:ultrastructure; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000648:ultrastructure; D015010:Yersinia pestis / Q000276:immunology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; C034284:IgG Fc-binding protein, Streptococcus; D010454:Peptide Termination Factors; D011993:Recombinant Fusion Proteins; C068068:SUP35 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C086725:caf1 protein, Yersinia pestis