Controlling biotinylation of microgels and modeling streptavidin uptake
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Compared are two approaches for the biotinylation of poly(N-isopropylacrylamide-co-vinylacetic acid) microgels, 300-nm diameter, water swollen particles with a corona of carboxyl groups. The biotinylated microgels are a platform for bioactive water-based ink. Streptavidin binding was measured as a function of biotin density, and the results were interpreted with a new model that predicts the minimum local density of biotins required to capture a streptavidin. An amino-polyethylene glycol derivative of biotin gave higher biotin contents than a biotin hydrazide. However, the streptavidin content versus biotin content results for both biotin derivatives fell on the same master curve with maximum biotin coverage of 0.11 mg of bound streptavidin per milligram of biotinylated microgel. Exclusion experiments showed that streptavidin was too big to penetrate the cross-linked microgel structure; thus, the conjugated streptavidin was restricted to the microgel surface. The colloidal stability of the microgels was preserved, and the biotinylated products showed good hydrolytic stability.
KeywordsMicrogel Biotinylation Bioconjugation Modeling
Prof. Todd Hoare is thanked for useful discussions. The authors acknowledge the SENTINEL Bioactive Paper NSERC Network for financial support. RP and YL hold Canada Research Chairs.
- 19.Hermanson GT (1996) Bioconjugate techniques. Academic, San DiegoGoogle Scholar
- 29.Kurzban GP, Bayer EA, Wilchek M, Horowitz PM (1991) The quaternary structure of streptavidin in urea. J Biol Chem 266(22):14470–14477Google Scholar