Abstract:
:Protein-based polymers are increasingly being used in biomaterial applications because of their ease of customization and potential monodispersity. These advantages make protein polymers excellent candidates for bioanalytical applications. Here we describe improved methods for producing drag-tags for free-solution conjugate electrophoresis (FSCE). FSCE utilizes a pure, monodisperse recombinant protein, tethered end-on to a ssDNA molecule, to enable DNA size separation in aqueous buffer. FSCE also provides a highly sensitive method to evaluate the polydispersity of a protein drag-tag and thus its suitability for bioanalytical uses. This method is able to detect slight differences in drag-tag charge or mass. We have devised an improved cloning, expression, and purification strategy that enables us to generate, for the first time, a truly monodisperse 20 kDa protein polymer and a nearly monodisperse 38 kDa protein. These newly produced proteins can be used as drag-tags to enable longer read DNA sequencing by free-solution microchannel electrophoresis.
journal_name
Biomacromoleculesjournal_title
Biomacromoleculesauthors
Lin JS,Albrecht JC,Meagher RJ,Wang X,Barron AEdoi
10.1021/bm200358rsubject
Has Abstractpub_date
2011-06-13 00:00:00pages
2275-84issue
6eissn
1525-7797issn
1526-4602journal_volume
12pub_type
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