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.

[1]  H. Sakaue,et al.  Climbing Rates of Microtubules Propelled by Dynein after Collision with Microfabricated Walls , 2012 .

[2]  B. Mowery,et al.  The paired t-test. , 2011, Pediatric nursing.

[3]  Siva K. Nalabotu,et al.  Transport of single cells using an actin bundle–myosin bionanomotor transport system , 2011, Nanotechnology.

[4]  Shimon Weiss,et al.  Aromatic aldehyde and hydrazine activated peptide coated quantum dots for easy bioconjugation and live cell imaging. , 2011, Bioconjugate chemistry.

[5]  A. Månsson,et al.  Long-Term Storage of Surface-Adsorbed Protein Machines , 2011, Langmuir : the ACS journal of surfaces and colloids.

[6]  Jennelle L. Malcos,et al.  Engineering tubulin: microtubule functionalization approaches for nanoscale device applications , 2011, Applied Microbiology and Biotechnology.

[7]  P. Walde,et al.  Preparation of catalytically active, covalent α-polylysine-enzyme conjugates via UV/vis-quantifiable bis-aryl hydrazone bond formation. , 2011, Biomacromolecules.

[8]  Ryan C Bailey,et al.  Efficient bioconjugation of protein capture agents to biosensor surfaces using aniline-catalyzed hydrazone ligation. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[9]  Edward H. Egelman,et al.  Structural Polymorphism in F-actin , 2010, Nature Structural &Molecular Biology.

[10]  Stefan Diez,et al.  Towards the application of cytoskeletal motor proteins in molecular detection and diagnostic devices. , 2010, Current opinion in biotechnology.

[11]  Miklós Nyitrai,et al.  Conformational Dynamics of Actin: Effectors and Implications for Biological Function , 2010, Cytoskeleton.

[12]  Leonid Ionov,et al.  Heavy meromyosin molecules extending more than 50 nm above adsorbing electronegative surfaces. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[13]  David M. Rissin,et al.  Single-Molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations , 2010, Nature Biotechnology.

[14]  Aaron R Wheeler,et al.  Immunoassays in microfluidic systems , 2010, Analytical and bioanalytical chemistry.

[15]  K. Kohama,et al.  Utilization of myosin and actin bundles for the transport of molecular cargo. , 2010, Small.

[16]  J. McDevitt,et al.  Programmable nano-bio-chip sensors: analytical meets clinical. , 2010, Analytical chemistry.

[17]  Amanda Carroll-Portillo,et al.  Directed attachment of antibodies to kinesin‐powered molecular shuttles , 2009, Biotechnology and bioengineering.

[18]  Chad A. Mirkin,et al.  Drivers of biodiagnostic development , 2009, Nature.

[19]  Amanda Carroll-Portillo,et al.  In vitro capture, transport, and detection of protein analytes using kinesin-based nanoharvesters. , 2009, Small.

[20]  S. Takeuchi,et al.  Biomolecular-motor-based nano- or microscale particle translocations on DNA microarrays. , 2009, Nano letters.

[21]  Ashutosh Agarwal,et al.  A smart dust biosensor powered by kinesin motors. , 2009, Nature nanotechnology.

[22]  Alf Månsson,et al.  Bending flexibility of actin filaments during motor-induced sliding. , 2008, Biophysical journal.

[23]  Lars Montelius,et al.  Diffusion dynamics of motor-driven transport: gradient production and self-organization of surfaces. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[24]  S. Diez,et al.  Setting up roadblocks for kinesin-1: mechanism for the selective speed control of cargo carrying microtubules. , 2008, Lab on a chip.

[25]  Takahiro Nitta,et al.  Comparing guiding track requirements for myosin- and kinesin-powered molecular shuttles. , 2008, Nano letters.

[26]  Banahalli R Ratna,et al.  Toward single molecule detection of staphylococcal enterotoxin B: mobile sandwich immunoassay on gliding microtubules. , 2008, Analytical chemistry.

[27]  Katsuo Kurabayashi,et al.  Self-contained, biomolecular motor-driven protein sorting and concentrating in an ultrasensitive microfluidic chip. , 2008, Nano letters.

[28]  William O Hancock,et al.  Transport and detection of unlabeled nucleotide targets by microtubules functionalized with molecular beacons , 2008, Biotechnology and bioengineering.

[29]  Viola Vogel,et al.  Cargo pick-up from engineered loading stations by kinesin driven molecular shuttles. , 2007, Lab on a chip.

[30]  A. Månsson,et al.  Contact angle measurements by confocal microscopy for non-destructive microscale surface characterization. , 2007, Journal of colloid and interface science.

[31]  Min-Gon Kim,et al.  Selective assembly and guiding of actomyosin using carbon nanotube network monolayer patterns. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[32]  Tai Kubo,et al.  Selective detection and transport of fully matched DNA by DNA-loaded microtubule and kinesin motor protein. , 2006, Biotechnology and Bioengineering.

[33]  G. Whitesides The origins and the future of microfluidics , 2006, Nature.

[34]  Lars Montelius,et al.  Actin filament guidance on a chip: toward high-throughput assays and lab-on-a-chip applications. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[35]  Lars Montelius,et al.  Selective spatial localization of actomyosin motor function by chemical surface patterning. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[36]  J. Beausang,et al.  Twirling of actin by myosins II and V observed via polarized TIRF in a modified gliding assay. , 2006, Biophysical journal.

[37]  Amanda Carroll-Portillo,et al.  Active capture and transport of virus particles using a biomolecular motor-driven, nanoscale antibody sandwich assay. , 2006, Small.

[38]  Viola Vogel,et al.  Selective loading of kinesin-powered molecular shuttles with protein cargo and its application to biosensing. , 2006, Small.

[39]  Toshio Yanagida,et al.  Dynamic polymorphism of single actin molecules in the actin filament , 2006, Nature chemical biology.

[40]  Peng Xiong,et al.  Packaging actomyosin-based biomolecular motor-driven devices for nanoactuator applications , 2005, IEEE Transactions on Advanced Packaging.

[41]  L. Montelius,et al.  Actin-Based Molecular Motors for Cargo Transportation in Nanotechnology— Potentials and Challenges , 2005, IEEE Transactions on Advanced Packaging.

[42]  H. Yeh,et al.  Single-quantum-dot-based DNA nanosensor , 2005, Nature materials.

[43]  Gengfeng Zheng,et al.  Multiplexed electrical detection of cancer markers with nanowire sensor arrays , 2005, Nature Biotechnology.

[44]  Lars Montelius,et al.  Guiding motor-propelled molecules with nanoscale precision through silanized bi-channel structures , 2005 .

[45]  Alf Månsson,et al.  Detection of small differences in actomyosin function using actin labeled with different phalloidin conjugates. , 2005, Analytical biochemistry.

[46]  C. Mirkin,et al.  Nanoparticle-based detection in cerebral spinal fluid of a soluble pathogenic biomarker for Alzheimer's disease. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[47]  Chad A. Mirkin,et al.  The use of nanoarrays for highly sensitive and selective detection of human immunodeficiency virus type 1 in plasma , 2004 .

[48]  M. Chee,et al.  Efficient strategies for the conjugation of oligonucleotides to antibodies enabling highly sensitive protein detection. , 2004, Biopolymers.

[49]  Lars Montelius,et al.  In vitro sliding of actin filaments labelled with single quantum dots. , 2004, Biochemical and biophysical research communications.

[50]  Lars Montelius,et al.  Silanized surfaces for in vitro studies of actomyosin function and nanotechnology applications. , 2003, Analytical biochemistry.

[51]  C. Mirkin,et al.  Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins , 2003, Science.

[52]  A. Månsson,et al.  Multivariate statistics in analysis of data from the in vitro motility assay. , 2003, Analytical biochemistry.

[53]  Viola Vogel,et al.  Light-Controlled Molecular Shuttles Made from Motor Proteins Carrying Cargo on Engineered Surfaces , 2001 .

[54]  D V Nicolau,et al.  Actin motion on microlithographically functionalized myosin surfaces and tracks. , 1999, Biophysical journal.

[55]  I. Sase,et al.  Axial rotation of sliding actin filaments revealed by single-fluorophore imaging. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[56]  S. Mashiko,et al.  Control of actin moving trajectory by patterned poly(methylmethacrylate) tracks. , 1997, Biophysical journal.

[57]  D. Murphy,et al.  Kinesin movement on glutaraldehyde-fixed microtubules. , 1996, Analytical biochemistry.

[58]  Iain D. C. Fraser,et al.  A simple method for automatic tracking of actin filaments in the motility assay , 1996, Journal of Muscle Research & Cell Motility.

[59]  M. Heidecker,et al.  Proximity relationships and structural dynamics of the phalloidin binding site of actin filaments in solution and on single actin filaments on heavy meromyosin. , 1995, Biochemistry.

[60]  E. Egelman,et al.  Structural dynamics of F-actin: II. Cooperativity in structural transitions. , 1995, Journal of molecular biology.

[61]  K C Holmes,et al.  Refinement of the F-actin model against X-ray fiber diffraction data by the use of a directed mutation algorithm. , 1993, Journal of molecular biology.

[62]  T. Yanagida,et al.  Inhibition of sliding movement of F-actin by crosslinking emphasizes the role of actin structure in the mechanism of motility. , 1990, Journal of molecular biology.

[63]  J. Spudich,et al.  Myosin step size. Estimation from slow sliding movement of actin over low densities of heavy meromyosin. , 1990, Journal of molecular biology.

[64]  W. Drabikowski,et al.  The effect of cytochalasin and glutaraldehyde on F-actin filaments containing muscle and non-muscle tropomyosin , 1983, Journal of Muscle Research & Cell Motility.

[65]  P. Sheehan,et al.  Attomolar protein detection in complex sample matrices with semi-homogeneous fluidic force discrimination assays. , 2009, Biosensors & bioelectronics.

[66]  A. Arner,et al.  Cardiotonic bipyridine amrinone slows myosin-induced actin filament sliding at saturating [MgATP] , 2004, Journal of Muscle Research & Cell Motility.

[67]  Z. Huang,et al.  Phallotoxin and actin binding assay by fluorescence enhancement. , 1992, Analytical biochemistry.

[68]  J. Spudich,et al.  Assays for actin sliding movement over myosin-coated surfaces. , 1991, Methods in enzymology.

[69]  J. Spudich,et al.  Purification of muscle actin. , 1982, Methods in enzymology.

[70]  James A. Spudich,et al.  Chapter 18 Purification of Muscle Actin , 1982 .

[71]  J. D. Pardee,et al.  [18] Purification of muscle actin , 1982 .