Antibodies covalently immobilized on actin filaments for fast myosin driven analyte transport.
Biosensors would benefit from further miniaturization, increased detection rate and independence from external pumps and other bulky equipment. Whereas transportation systems built around molecular motors and cytoskeletal filaments hold significant promise in the latter regard, recent proof-of-principle devices based on the microtubule-kinesin motor system have not matched the speed of existing methods. An attractive solution to overcome this limitation would be the use of myosin driven propulsion of actin filaments which offers motility one order of magnitude faster than the kinesin-microtubule system. Here, we realized a necessary requirement for the use of the actomyosin system in biosensing devices, namely covalent attachment of antibodies to actin filaments using heterobifunctional cross-linkers. We also demonstrated consistent and rapid myosin II driven transport where velocity and the fraction of motile actin filaments was negligibly affected by the presence of antibody-antigen complexes at rather high density (>20 µm(-1)). The results, however, also demonstrated that it was challenging to consistently achieve high density of functional antibodies along the actin filament, and optimization of the covalent coupling procedure to increase labeling density should be a major focus for future work. Despite the remaining challenges, the reported advances are important steps towards considerably faster nanoseparation than shown for previous molecular motor based devices, and enhanced miniaturization because of high bending flexibility of actin filaments.
School of Natural Sciences, Linnaeus University, Kalmar, Sweden.
Saroj
S
Kumar
Lasse
L
ten Siethoff
Malin
M
Persson
Mercy
M
Lard
Geertruy
G
te Kronnie
Heiner
H
Linke
Alf
A
Månsson
D000199:Actins / Q000276:immunology*; D000906:Antibodies / Q000276:immunology*; D015374:Biosensing Techniques; D008856:Microscopy, Fluorescence; D009218:Myosins / Q000276:immunology*; D013056:Spectrophotometry, Ultraviolet
D000199:Actins; D000906:Antibodies; D009218:Myosins