Team:LMU-TUM Munich/Linkerchemistry

Kopiervorlagen

Seitenverantwortliche/r:Vivien

Bei Google Scholar bitte das APA-Ziteirformat verwenden.

Textformatierung

kursiv
fett
Strich

Links

Wikiinterner Link Team:LMU-TUM_Munich/Materials (As described in the Materials section)

Wikiexterner Link Visit W3Schools
Visit W3Schools

Bilder

Bildunterschrift





















Introduction

Purpose

Any cell mesh produced based on our bioink will suffer from exposition to serum-containing media or, in the case of in vivo application, the patient’s body fluids because of biotinidase. This ubiquitous enzyme is able to cleave biotin off of lysine – at a lower rate also off of intact protein. We may now shift this hydrolysis in two directions: either to prevent it altogether and thus stabilize our cell mesh, or to accelerate it to make our mesh quickly disintegrable. To this end, we must consider how biotin is linked to the proteins in our bioink.

NHS Esters as Amine Coupling Reagents

N-hydroxysuccinimide (NHS) esters belong to the group of amine-reactive active esters and are thus of great use in labeling proteins. Typically, NHS esters react with the primary amino group in the side chain of surficial lysine, forming a stable peptide bond.[1]

The utility of NHS esters was first suggested by Anderson et al.[2] in the context of peptide synthesis; nowadays, a plethora of NHS esters are widely used in bioconjugate techniques and thus commercially available, including those of many fluorescent dyes as well as that of biotin, often referred to as biotin-NHS (Figure 1).[3] NHS esters are typically synthesized from the corresponding carboxylic ester and NHS via the one-pot DCC coupling procedure [1] and purified by crystallization[2][3]
  1. ↑ 1.0 1.1 Hermanson, G. T. (2013) Bioconjugate Techniques (3 ed.). Boston: Academic Press.
  2. ↑ 2.0 2.1 Anderson, G. W., Zimmerman, J. E., & Callahan, F. M. (1963). N-Hydroxysuccinimide Esters in Peptide Synthesis. Journal of the American Chemical Society, 85(19), 3039-3039.
  3. ↑ 3.0 3.1 Klykov, O., & Weller, M. G. (2015). Quantification of N-hydroxysuccinimide and N-hydroxysulfosuccinimide by hydrophilic interaction chromatography (HILIC). Analytical Methods, 7(15), 6443-6448.