While the order of mixing shows a prominent effect at lower stoichiometries of mixing, higher NaCl salt reduces the effect all together. DLS will generally measure a wider size range than NTA, but NTA offers greater resolution than DLS (even with Multi-angle Dynamic Light Scattering). A Malvern Instruments Bioscience Development Initiative Executive Summary. Routes to control the complexation regimes with elevated NaCl or KCl salt concentrations indicate ion-specific effects, such that more smaller-size complexes are present at higher NaCl, counterintuitive with respect to PCPP solubility arguments. 15 Apr, 2019 Particle Science - Guide Nanoparticle Tracking Analysis (NTA) and Dynamic Light Scattering (DLS) are complementary techniques that offer different insights into your samples. The antigenicity of PCPP bound lysozyme, when compared to equivalent lysozyme solutions, was largely retained for all complexes, but observed a dramatic reduction for heavily aggregated systems. Three regimes of self-assembly were observed covering complexation of PCPP with lysozyme in the nano-scale range, multi-chain complexes, and larger aggregates with complexes characterized by a maximum loading of over six hundred protein molecules per PCPP chain and dissociation constant in the micromolar range (Kd = 7 × 10–6 mol/L). In the scope of DLS, temporal fluctuations are usually analyzed using the intensity or photon auto-correlation function (also known as photon correlation spectroscopy or. The formation of supramolecular complexes between polyphosphazene adjuvant, poly (PCPP), and a model vaccine antigen, hen egg lysozyme, was studied under physiological conditions using automated dynamic light scattering titration, asymmetric flow field flow fractionation (AF4), enzyme-linked immunosorbent assay (ELISA), and fluorescent quenching methods. Dynamic light scattering (DLS) is a technique in physics that can be used to determine the size distribution profile of small particles in suspension or polymers in solution. In this third white paper of the series, we cover the basic types of DLS deconvolution algorithms used to extract the intensity weighted particle size distribution from the measured correlogram.Polyphosphazenes represent a class of intrinsically flexible polyelectrolytes with potent immunoadjuvant activity, which is enabled through non-covalent self-assembly with antigenic proteins by charge complexation. Concentration Effects and Particle Interactions 07 February 2014 Contact sales The influence of concentration effects and particle interactions on DLS results is discussed at the example of increasing sample concentration. plasma proteins. Application of Dynamic Light Scattering (DLS) to Protein Therapeutic Formulations: Principles, Measurements and Analysis - 2. In this four-part white paper series, common issues and questions surrounding the principles, measurements and analysis of DLS data are discussed in order to help minimize the time required for and complexity of acquiring and interpreting DLS data that is critical throughout the development process. properties such as hydrodynamic dimensions can potentially be extracted from dynamic light scattering (DLS) measurements on SWCNTs done in the solution state, however. With that said, the challenge that remains with the application of DLS to protein therapeutic formulations is centered around data interpretation. The popularity of DLS within the biopharmaceutical industry is a consequence of its wide working size and extended sample concentration ranges, as well as its low volume requirements. A Malvern Instruments' Bioscience Development Initiativeĭynamic light scattering (DLS) is an analytical technique used to measure the particle size distribution of protein formulations across the oligomer and sub-micron size ranges of approximately 1 nm to 1 µm.
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