Directed attachment of antibodies to kinesin-powered molecular shuttles.
Biomolecular motors, such as kinesin, have been used to shuttle a range of biological and synthetic cargo in microfluidic architectures. A critical gap in this technology is the ability to controllably link macromolecular cargo on microtubule (MT) shuttles without forming extraneous byproducts that may potentially limit their application. Here we present a generalized approach for functionalizing MTs with antibodies in which covalent bonds are formed between the carbohydrate in F(c) region of polyclonal antibodies and the positively charged amino acids on the MT surface using the crosslinker succinimidyl 4-hydrazidoterephthalate hydrochloride (SHTH). Antibody-functionalized MTs (Ab-MTs) produced through this approach maintained motility characteristics and antigenic selectivity, and did not produce undesirable byproducts common to other approaches. We also demonstrate and characterize the application of these Ab-MTs for capturing and transporting bacterial and viral antigens. While this approach cannot be applied to monoclonal antibodies, which lack a carbohydrate moiety, it may be used for selectively functionalizing MT shuttles with a variety of carbohydrate-containing cargoes.
Carroll-Portillo
Amanda
A
Center for Integrated Nanotechnologies, Albuquerque, New Mexico 87185, USA.
Bachand
Marlene
M
Bachand
George D
GD
D000906:Antibodies / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000378:metabolism; D016547:Kinesins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D011485:Protein Binding
D000906:Antibodies; D003432:Cross-Linking Reagents; D016547:Kinesins