Efficiency optimisation of proteins on a chip.
This study elucidates that the protein reorientation on a chip can be changed by an external electric field (EEF) and optimised for achieving strong effective binding between proteins. Protein A and its binding protein immunoglobulin G (IgG) were used as an example, in addition to an anticancer peptide (CB1a) and its antibody (anti-CB1a). The binding forces (BFs) were measured by atomic force microscopy (AFM) with EEFs applied at different angles (EEF°). The optimal angle (OA) of the EEF (OAEEF°) corresponding to the maximum binding force (BFmax) was obtained. The results showed that the BFmax values between IgG/Protein A and anti-CB1a/CB1a were 6424.2 ± 195.3 pN (OAEEF° = 45°) and 729.1 ± 33.2 pN (OAEEF° = 22.5°), respectively. Without an EEF, the BF values were only 730.0 ± 113.9 pN and 337.3 ± 35.0 pN, respectively. Based on these observations, we concluded that the efficient optimisation of protein-protein interaction on a chip is essential. This finding is applicable to the industrial fabrication of all protein chips.
National Nano Device Laboratories, National Applied Research Laboratories, Hsinchu 300, Taiwan, ROC. hmchen@narlabs.org.tw.
Wei-jen
WJ
Wu
Hsuan-yu
HY
Huang
Wei-Yeh
WY
Hsu
Ray-Quen
RQ
Hsu
Hueih-Min
HM
Chen
D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000937:Antigen-Antibody Reactions; D023181:Antimicrobial Cationic Peptides / Q000032:analysis / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D018625:Microscopy, Atomic Force; D040081:Protein Array Analysis; D013205:Staphylococcal Protein A / Q000276:immunology
D000906:Antibodies; D023181:Antimicrobial Cationic Peptides; C000598900:CB1a protein; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A