High-capacity binding of proteins by poly(acrylic acid) brushes and their derivatives

Langmuir : the ACS Journal of Surfaces and Colloids
Jinhua DaiMerlin L Bruening

Abstract

Polymeric coatings with high protein-binding capacities are important for increasing the output of affinity-based protein purification and decreasing the detection limits of antibody microarrays. This report describes the use of thick poly(acrylic acid) (PAA) brushes to immobilize as much as 80 monolayers of protein. The brushes were prepared using a recently developed procedure that allows polymerization of 100-nm-thick poly(tert-butyl acrylate) films from a surface in just 5 min along with hydrolysis of these films to PAA in 15 min. Covalent binding of bovine serum albumin (BSA) to PAA brushes that were activated using standard coupling agents, however, resulted in immobilization of less than two monolayers of BSA because of competitive hydrolysis of the esters in the activated film. In contrast, derivatization of PAA with nitrilotriacetate (NTA)-Cu2+ complexes yielded films capable of binding many monolayers of protein via metal-ion affinity interactions. For example, derivatization of 55-nm-thick PAA films with NTA-Cu2+ allowed immobilization of about 15 monolayers (5.8 microg/cm2 or 58 nm) of BSA. The binding capacity was even higher for myoglobin (7.7 microg/cm2) and anti-IgG (9.6 microg/cm2). Remarkably, electrostatic ad...Continue Reading

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Citations

Jan 31, 2013·ACS Applied Materials & Interfaces·G Wilhelmina de GrootG Julius Vancso
May 12, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Ethan N ChiangBarbara A Baird
Mar 24, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Nicolas Schüwer, Harm-Anton Klok
Aug 23, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Cuie WangShou-Jun Xiao
Apr 4, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Somnath BhattacharjeeMerlin L Bruening
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Dec 24, 2015·Langmuir : the ACS Journal of Surfaces and Colloids·Oraphan WiarachaiVoravee P Hoven
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