Mempro™ Chromatography Services

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With the well-established Mempro™ nanodiscs analysis platform, Creative Biostructure provides customized Mempro™ nanodisc analysis service using chromatography technique. Our highly professional scientists are confident in providing chromatography service and offering the best service to the customers around the world.

Chromatography is a useful technique for isolating the solutes or components from mixtures based on the relative numbers of each solute distributed between a flowing fluid stream, namely the mobile phase, and an adjoining changeless phase. The mobile phase can be either a gas or a liquid, and the stationary phase is either a liquid or a solid. Solute molecules are successively swapped through kinetic molecular motion between the two phases. Nanodiscs, soluble nanoscale lipid bilayers, are preeminent biomimetic systems for researching the membrane-anchored and entire membrane protein complexes under certain experimental states. The capacity to acquire samples of a restrained size distribution is an important feature of nanodiscs, and the usual method of the size-dependent nanodisc purification after reconstitution is size exclusion chromatography (SEC).

Figure 1. A schematic of the SPR sensor chips and their capture strategies.( Trahey, M. 2015)

Nanodiscs Analysis Using Size Exclusion Chromatography (SEC)
The SEC technique has been applied for the beginning of nanodisc reconstitution and keeps a nearly mandatory step in the nanodisc sample preparation. SEC is extremely suitable to remove any aggregates from nanodisc reconstitution and for the evaluation of the dimensions and homogeneity of the samples. Most published work on nanodiscs related to SEC uses a Superdex 200 column, which is well appropriate for purifying and analyzing nanodiscs. Moreover, dynamic light scattering has been applied for determining the molecular weight and dimension of dimension through combining with SEC.

SEC is also an available tool for the identification of right nanodisc assembly and membrane protein reconstitution. Generally, a reinjection of portions covering the nanodisc summit is performed to verify the stability of the reconstituted samples. The homogeneity of the nanodiscs is confirmed from the appearance of a unique elution peak, and membrane protein reconstitution is measured from the coevolution of the MSPs and the target membrane protein. This is usually analyzed by the sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), or another method that uses fluorescent-labeled membrane proteins, which make the absorption detectable immediately in the chromatogram.

Creative Biostructure has been focusing on nanodiscs technology for a long time, with the comprehensive nanodisc analysis platform, we can perform the incomparable chromatography services for customers. Besides nanodiscs analysis, Creative Biostructure also provides other various Mempro™ nanodiscs technology services. Please feel free to contact us for a detailed quote.

References:
Trahey, M. (2015). “Applications of Lipid Nanodiscs for the Study of Membrane Proteins by Surface Plasmon Resonance”. Current Protocols in Protein Science, 29-13.
Debu, D. T. (2016). “Effect of Ti adhesive layer on individual gold nanodisk surface plasmon resonance”. In APS Meeting Abstracts.