Functionalization of Magnetic Beads with Lipobiotin

5.6 x 10⁸ magnetic beads (Micromer-M, polystyrene body, surface: PEG-COOH, 5μm; in dH₂0) were incubated with 12.8 mg N-hydroxysuccinimide (NHS) and 6.4 mg 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) in 20% [v/v] 2-(N-morpholino)ethanesulfonic acid (MES) buffer (pH 6.3) and agitated for 1.5 hours. Activated beads were washed twice with PBS and then incubated with the biotinylated lipopeptide lipobiotin (PHCKKKKK(Aca-Aca-Biotin) x 3 TFA, N-Palmitoyl-S-(1,2-bishexadecyloxy-carbonyl) ethyl-[R]-cysteinyl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]-lysine(ɛ-aminocaproyl-ϵ-aminocaproyl-biotinyl) x 3 CF3COOH (EMC Microcollections, Tübingen, Germany) at 1.5 mg/ml in carbonate buffer (pH 9.2) at RT for 4.5 hours. Functionalized beads were washed twice with PBS and incubated in 10mM ethanolamine (in carbonate buffer; pH 9.2) to block unreacted ester groups (1h, RT). Lipobiotin-functionalized magnetic beads (LMB) were washed, re-suspended in PBS and kept at 4°C. Non-functionalized magnetic beads (NMB) that were equally washed and re-suspended in PBS served as a control.

Free full text: Click here

Hansen J., Kolbe K., König I.R., Scherließ R., Hellfritzsch M., Malm S., Müller-Loennies S., Zallet J., Hillemann D., Wiesmüller K.H., Herzmann C., Brandenburg J, & Reiling N. (2022). Lipobiotin-capture magnetic bead assay for isolation, enrichment and detection of Mycobacterium tuberculosis from saliva. PLoS ONE, 17(7), e0265554.

Publication 2022

Biotin Block Body Buffer Carbodiimide Carbonate Ester Ethanesulfonic acid Ethanolamine Lipopeptide Lysine Morpholino Polystyrene

Corresponding Organization : German Center for Infection Research

Other organizations : University of Lübeck, University Hospital Schleswig-Holstein, Kiel University, EMC Microcollections (Germany)

Top 5 similar protocols

Variable analysis

independent variables

Concentration of N-hydroxysuccinimide (NHS)
Concentration of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)
Concentration of biotinylated lipopeptide lipobiotin
Incubation time with NHS and EDC
Incubation time with biotinylated lipopeptide lipobiotin

dependent variables

Binding of biotinylated lipopeptide lipobiotin to magnetic beads

control variables

Concentration of magnetic beads (5.6 x 10^8 beads)
Bead characteristics (Micromer-M, polystyrene body, surface: PEG-COOH, 5μm)
Incubation buffer conditions (20% [v/v] 2-(N-morpholino)ethanesulfonic acid (MES) buffer, pH 6.3; carbonate buffer, pH 9.2)
Washing steps (twice with PBS)

positive control

Lipobiotin-functionalized magnetic beads (LMB)

negative control

Non-functionalized magnetic beads (NMB)

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!