By Vladimir V. Shuvaev, Thomas Dziubla, Rainer Wiewrodt, Vladimir R. Muzykantov (auth.), Christof M. Niemeyer (eds.)
Professional laboratorians replace the vintage bioconjugation tools and introduce worthy new ideas that transcend natural biomedical functions to incorporate parts from complex natural synthesis, molecular biology, and fabrics technological know-how. those effectively reproducible tools disguise the education of protein conjugates utilizing covalent and noncovalent conjugation, the synthesis of nucleic acid conjugates utilizing various labeling thoughts, and methods to semisynthetic conjugates of proteins. extra chapters tackle the biofunctionalization of inorganic surfaces, together with the on-chip synthesis of peptide nucleic acids to generate microarrays for the high-throughput research of RNA and DNA, gold nanaoparticles, and carbon nanotube probes for atomic strength microscopy.
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Additional info for Bioconjugation Protocols: Strategies and Methods
263, 18,220–18,224. METHODS IN MOLECULAR BIOLOGY TM TM Volume 283 Bioconjugation Protocols Strategies and Methods Edited by Christof M. Niemeyer Smart Polymer–Streptavidin Conjugates 37 3 Smart Polymer–Streptavidin Conjugates Patrick S. Stayton, Zhongli Ding, and Allan S. Hoffman Summary The conjugation of stimuli-responsive, or “smart,” polymers to streptavidin is described. The polymer is synthesized with a thiol-reactive end-group, which is used to end-graft the polymer to cysteine or lysine side-chains that are genetically engineered into controlled positions on the streptavidin surface.
2. COUPLING WITH PEG–VS 1. 0 (either borate or tris) containing 1–5 mM EDTA as a scavenger. 0 because an amine reaction can occur (even if slow). If you intend to follow the reaction by thiol titration using the Ellman’s assay, keep aside a small portion of protein solution as unmodified protein reference solution (N). 2. Add PEG–VS. The VS group is less reactive than maleimide. To accelerate the reaction, more than one equivalent (2–10) of reagent for each thiol may be used (see Table 1). 3. 5–8 h, depending on the amount of PEG reagent used.
S. (1995) Site-directed mutagenesis studies of the high affinity streptavidin-biotin complex: contributions of tryptophan residues 79, 108, and 120. Proc. Natl. Acad. Sci. USA 92, 1754–1758. 9. Riddles, P. , Blakeley, R. L. and Zerner, B. (1983) Reassessment of Ellman’s reagent. Methods Enzymol. 91, 49–60. METHODS IN MOLECULAR BIOLOGY TM TM Volume 283 Bioconjugation Protocols Strategies and Methods Edited by Christof M. Niemeyer Conjugates of Peptides and Proteins 45 4 Conjugates of Peptides and Proteins to Polyethylene Glycols Margherita Morpurgo and Francesco M.