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Directed attachment of antibodies to kinesin‐powered molecular shuttles
Author(s) -
CarrollPortillo Amanda,
Bachand Marlene,
Bachand George D.
Publication year - 2009
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22501
Subject(s) - kinesin , polyclonal antibodies , monoclonal antibody , chemistry , covalent bond , microtubule , biochemistry , antibody , macromolecule , antigen , protein engineering , biophysics , combinatorial chemistry , biology , microbiology and biotechnology , enzyme , organic chemistry , immunology , genetics
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. Biotechnol. Bioeng. 2009; 104: 1182–1188. © 2009 Wiley Periodicals, Inc.