Understanding the colloidal stability of protein therapeutics using dynamic light scattering

Size: px
Start display at page:

Download "Understanding the colloidal stability of protein therapeutics using dynamic light scattering"

Transcription

1 Understanding the colloidal stability of protein therapeutics using dynamic light scattering A Malvern Instruments' Bioscience Development Initiative Executive summary The combination of dynamic light scattering (DLS) with Raman spectroscopy provides the ability to extract a wealth of chemical, structural, and physical information about biotherapeutic proteins under formulation conditions. Dynamic light scattering (DLS) using noninvasive backscatter (NIBS) detection technology is capable of measuring the hydrodynamic radius of proteins over a large concentration range (0.1 mg/ml to 100 mg/ml). As a result, it is possible to determine, in actual biopharmaceutical formulations, the bulk viscosity/restricted diffusion interaction parameter (k D ), particle interaction parameter (B 22 ), melting temperature, onset temperature of protein aggregation and transition enthalpies to assess colloidal stability. The unique coupling of DLS with Raman spectroscopy provides the ability to simultaneously correlate colloidal parameters to protein structure under a variety of stress conditions, e.g., thermal, formulation, chemical degradation, extrinsic particulates, to enhance our understanding of protein therapeutic formulations. Introduction Dynamic light scattering (DLS) is a workhorse technique for determining the hydrodynamic diameter of biotherapeutic proteins in solution, sample polydispersity, and sample interactions. Raman spectroscopy derives information about protein secondary and tertiary structure by monitoring molecular vibrations. By combining these two analytical approaches into a single system, a wealth of chemical, structural and physical parameters can be determined. Protein secondary and tertiary structure, melting temperature, onset temperature of aggregation and transition enthalpy values can all be derived, as well as aggregation propensity, protein solubility, and the potential for high viscosities at formulated concentrations. The complementary nature of the results obtained from DLS and Raman spectroscopy on the same sample may provide unique insights into the mechanisms of aggregation and unfolding. Here we Malvern Instruments Worldwide Sales and service centres in over 65 countries Malvern Instruments Limited

2 describe the utility of DLS to aid in the understanding the colloidal stability of protein therapeutics. The approach Malvern Instruments Zetasizer Helix (ZS Helix) integrates a fiber-coupled Raman spectrometer with a Zetasizer Nano ZSP to provide DLS (colloidal stability) and Raman (conformational stability) data sequentially on a single sample. The Zetasizer Nano system integrates proprietary non-invasive backscatter (NIBS) detector technology with dynamic (DLS), static (SLS) and electrophoretic (ELS) light scattering to measure the hydrodynamic radius of proteins from 0.15 nm - 5 µm, from 0.1 mg/ ml to 100 mg/ml. Raman spectra are collected using 785 nm excitation (~280 mw) from cm -1 at 4 cm -1 resolution. Sample aliquots (~120 µl) are placed into a 3 mm quartz cuvette and positioned in a temperature-controlled compartment that provides temperature control from 0 C - 90 C ± 0.1 C. Thermal ramp studies are conducted by collecting Raman and DLS data over a series of pre-defined 0.1 C - 5 C step-wise increments. Isothermal incubation studies are conducted by collecting a series of Raman and DLS data over a predefined time interval at a desired temperature set-point. Results and Discussion Measuring protein size at high concentration The formulation of monoclonal antibodies at high concentration has drawn great attention within the biopharmaceutical industry in recent years. However, for DLS, size measurement at formulated concentrations was limited by multiple experimental factors. As shown in Figure 1, increasing sample concentration could affect the observed scattering intensity and consequently the size determination. Factors affecting the measurement could include: multiple scattering, viscosity, positive/ negative virial effects, and self-association. Deconvolution of the above effects allows for interpretation of high-concentration sample results. Figure 1: The observed effects of increased sample concentration on the measured light scattering intensity. 2 Understanding the colloidal stability of protein therapeutics using dynamic light scattering

3 To better understand the effects of the aforementioned factors on the DLS data, it is convenient to consider an individual effect in isolation. Table I (below) summarizes the expected changes with increasing sample concentration to the reported size, polydispersity, and correlation function, as well as established correction methods, for each of the unwanted factors. Table 1: Summary of the effects of high concentration sample on DLS parameters Factors Apparent Size Apparent Polydispersity Correlation Intersection Correction Method Multiple Scattering Decrease Increase Decrease Backscattering Restricted Diffusion Increase No Change No Change Viscosity Correction Negative B 22 Increase Increase No Change No Positive B 22 Decrease No Change No Change No Figure 2 (below) demonstrates the effect of viscosity correction on the reported size of a sample. Using the viscosity of water as an estimate of the sample viscosity results in an increase in apparent size of the suspended particles (blue squares). However, when the true bulk solution viscosity is used to correctly fit the DLS results, no apparent size change is observed (green circles). Particle interaction terms including attractive/ repulsive, B 22 (second virial coefficient) or k d (diffusion interaction parameter) could be indicated and will be covered in the following section. It should be noted that for virial effects, there is currently no agreed approach to correct DLS data. Figure 2: The effects of bulk viscosity correction on the size measurement results. Detecting protein aggregation onset Static light scattering (SLS) or turbidity coupled with thermal ramping have been routinely used as rapid formulation screening tools for protein stability. Essentially, these methods measure the aggregation onset temperature T onset. DLS could also be utilized in this fashion. Figure 3 shows the aggregation onset for BSA is approximately 62 C by Z-averaged size. Additionally, as long as the correlogram remains valid, DLS offers the size distribution (Fig. 3, insert) and polydispersity (PDI), where SLS offers only the intensity. The size distribution trend shown here clearly reveals the 3 Understanding the colloidal stability of protein therapeutics using dynamic light scattering

4 appearance of a second peak at 90 nm near 60 C. The onset temperature from PID could be as early as 55 C, indicating aggregation may begin even earlier. Figure 3: DLS thermal ramping of BSA (50 mg/ml ph 7.4). Figure 4: Illustration of the intensity profile of two particles with 5 and 50 nm diameter. The intensity ratio will be 1: The ability to detect trace amounts of aggregates stems from the inherent sensitivity of DLS. Figure 4 demonstrates that for a simple mixture of two particles with an order of magnitude difference in diameter, the larger 50 nm particle will show a scattering intensity a million times stronger than the smaller 5 nm particle. This infers that even a single large particle mixed with a million small particles results in an equal detected intensity! Thus, DLS could be useful as the earliest detector for the appearance of aggregates. However, it should be noted that the resolution of DLS is inherently limited to a factor of three in size. Predicting colloidal stability 4 Understanding the colloidal stability of protein therapeutics using dynamic light scattering

5 Though T onset might be sufficient for initial formulation screening, no understanding of protein interaction can be reached from such experiments, let alone the prediction of protein stability. Recently, more attention has been focused on investigating intermolecular parameters as predictors of protein aggregation kinetics, which have been successfully correlated with B 22 and k D [1]. A large positive value of B 22 indicates that solutes prefer association with the solvent over self-association, either due to strong electrostatic repulsion between solute molecules or positive interaction between solute and solvent. In contrast, large negative values of B 22 indicate strongly selfattractive systems, where the particles prefer self-association or aggregation over complete solvation. In the case of DLS, k D can be measured directly and is related to B 22 by the following equations [2]: Here, C is the sample concentration and D refers to the diffusion coefficient measured by DLS. D 0 is the self-diffusion coefficient (the value of D at zero concentration). M w is the sample molecular weight, k f is the first order concentration coefficient of the friction and ν is the partial specific volume of the solute. The following experiments demonstrate how to apply k D to predict the protein stability for an IgG in four formulations. In Figure 5 only Soup 2 (red circles) has a positive k D, indicating a strong repulsive interaction between the protein samples. The other three samples show slightly negative k D. It should be mentioned that large positive k D usually indicates strong repulsive interactions, or positive B 22. However, when k D is close to zero it becomes difficult to directly link to B 22. Figure 5: k D plot measured for the same IgG solution in four formulations. 5 Understanding the colloidal stability of protein therapeutics using dynamic light scattering

6 Figure 6: DLS thermal ramping of one IgG sample in four different formulations. SLS can be measured at a fixed angle for B 22, and zeta potential measured to provide relative charge, as summarized in Table 2. Putting all of the information together, a comprehensive profile of protein properties can be obtained to predict protein stability. For example, we would predict that formulation 2 is the most stable formulation for this IgG, while formulation 1 is the least stable. To support this prediction, thermal ramping experiments were performed on the same set of samples, and are shown in Figure 6. Interestingly, sample 1 has the lowest T onset, close to 66 C, while formulation 2 is the most stable, showing no evidence of large aggregates up to 90 C. Table 2: Summary of predictive parameters for IgG stability IgG Formulation B 22 (10-5 ml mol/g 2 ) k D (ml/g) Charge Soup Soup Soup Soup Summary The combination and integration of DLS and Raman spectroscopy into a single instrumental platform provides unique analytical capabilities to determine protein secondary and tertiary structure, melting temperature, and size and onset of aggregation for the same sample, without altering the testing conditions. Here we have described the unique ability of DLS, when combined with noninvasive backscatter detector (NIBS) technology, to derive the bulk viscosity/restricted diffusion interaction parameter (k D ), particle interaction parameter (B 22 ), melting temperature, onset temperature of aggregation and transition enthalpies on high concentration formulations. A subsequent study describes the unique spectra-structure properties that may be correlated using Raman Spectroscopy (Understanding the Conformational Stability of Protein Therapeutics with Raman Spectroscopy). Future studies will report on the unique correlations that may be derived from the combination of DLS and Raman spectroscopy to improve the understanding of the kinetics, thermodynamics 6 Understanding the colloidal stability of protein therapeutics using dynamic light scattering

7 and mechanism(s) of aggregation and the correlation of specific protein structural motifs to aggregation to improve product knowledge and formulation stability. About Malvern's Bioscience Development Initiative Malvern Instruments' Bioscience Development Initiative (BDI) was established to accelerate innovation, development and the promotion of new technologies, products and capabilities to address unmet measurement needs in the biosciences markets. References 1. Diffusion and sedimentation interaction parameters for measuring the second virial coefficient and their utility as predictors of protein aggregation. Saluja A. et al., Biophysical Journal , Specific interactions in high concentration antibody solutions resulting in high viscosity. Yadav S. et al., Journal of Pharm. Sci. 2010, 99, Understanding the colloidal stability of protein therapeutics using dynamic light scattering

8 Malvern Instruments Limited Grovewood Road, Malvern, Worcestershire, UK. WR14 1XZ Tel: Fax: Malvern Instruments is part of Spectris plc, the Precision Instrumentation and Controls Company. Spectris and the Spectris logo are Trade Marks of Spectris plc. All information supplied within is correct at time of publication. Malvern Instruments pursues a policy of continual improvement due to technical development. We therefore reserve the right to deviate from information, descriptions, and specifications in this publication without notice. Malvern Instruments shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing, performance or use of this material. Malvern Instruments owns the following registered trademarks: Bohlin, FIPA, Insitec, ISYS, Kinexus, Malvern, Malvern 'Hills' logo, Mastersizer, Morphologi, Rosand, 'SEC-MALS', Viscosizer, Viscotek, Viscogel and Zetasizer Malvern Instruments Limited

Use of DLS/Raman to study the thermal unfolding process of lysozyme

Use of DLS/Raman to study the thermal unfolding process of lysozyme Use of DLS/Raman to study the thermal unfolding process of lysozyme A Malvern Instruments' Bioscience Development Initiative Executive summary The combination of dynamic light scattering (DLS) and Raman

More information

Understanding the conformational stability of protein therapeutics using Raman spectroscopy

Understanding the conformational stability of protein therapeutics using Raman spectroscopy Understanding the conformational stability of protein therapeutics using Raman spectroscopy A Malvern Instruments' Bioscience Development Initiative Executive summary The combination of Dynamic Light Scattering

More information

DEVELOPING A BIOFORMULATION STABILITY PROFILE

DEVELOPING A BIOFORMULATION STABILITY PROFILE DEVELOPING A BIOFORMULATION STABILITY PROFILE Light Scattering and Micro-Capillary Viscometry A Malvern Instruments Bioscience Development Initiative Executive Summary: When used in combination, light

More information

Introduction to the calculators in the Zetasizer software

Introduction to the calculators in the Zetasizer software Introduction to the calculators in the Zetasizer software PARTICLE SIZE ZETA POTENTIAL MOLECULAR WEIGHT MOLECULAR SIZE Introduction The calculators are a series of tools in the Zetasizer software that

More information

The power of multi-detector SEC in the analysis of antibodies

The power of multi-detector SEC in the analysis of antibodies The power of multi-detector SEC in the analysis of antibodies How OMNISEC can measure antibody absolute MW independent of size and structural changes MOLECULAR SIZE MOLECULAR STRUCTURE MOLECULAR WEIGHT

More information

Executive Summary. Introduction WHITEPAPER

Executive Summary. Introduction WHITEPAPER The Characterization of Single Wall Carbon Nanotubes Using The Malvern Instruments Helix System: Integrating Dynamic Light Scattering (DLS) and Raman Spectroscopy Executive Summary The combination of DLS

More information

Component specific particle characterisation of the active components in a pharmaceutical topical formulations

Component specific particle characterisation of the active components in a pharmaceutical topical formulations Component specific particle characterisation of the active components in a pharmaceutical topical formulations PARTICLE SIZE PARTICLE SHAPE CHEMICAL IDENTIFICATION Introduction Most pharmaceutical products

More information

Characterization of Single Wall Carbon Nanotubes (SWNTs) by combining Dynamic Light Scattering and Raman Spectroscopy

Characterization of Single Wall Carbon Nanotubes (SWNTs) by combining Dynamic Light Scattering and Raman Spectroscopy Characterization of Single Wall Carbon Nanotubes (SWNTs) by combining Dynamic Light Scattering and Raman Spectroscopy PARTICLE SIZE Executive Summary The combination of DLS and Raman spectroscopy offers

More information

GPC/SEC Analysis of Polymer Degradation

GPC/SEC Analysis of Polymer Degradation MOLECULAR WEIGHT Introduction MOLECULAR Understanding the effects of processing a polymer on its molecular weight, STRUCTURE molecular weight distribution, and solution viscosity are important to MOLECULAR

More information

Discriminating between polymorphs of acetaminophen using Morphologically-Directed Raman Spectroscopy (MDRS)

Discriminating between polymorphs of acetaminophen using Morphologically-Directed Raman Spectroscopy (MDRS) Discriminating between polymorphs of acetaminophen using Morphologically-Directed Raman Spectroscopy (MDRS) CHEMICAL IDENTIFICATION PARTICLE SIZE PARTICLE SHAPE Introduction Interest in studying mixed

More information

Optimization of PLA/PLGA molecular weight and structure to suit final application requirements

Optimization of PLA/PLGA molecular weight and structure to suit final application requirements Optimization of PLA/PLGA molecular weight and structure to suit final application requirements MOLECULAR SIZE MOLECULAR STRUCTURE MOLECULAR WEIGHT Introduction The biodegradable and biocompatible polymers,

More information

Characterization of protein aggregates in suspension and on a filter membrane by Morphologically-Directed Raman Spectroscopy

Characterization of protein aggregates in suspension and on a filter membrane by Morphologically-Directed Raman Spectroscopy Characterization of protein aggregates in suspension and on a filter membrane by Morphologically-Directed Raman Spectroscopy Morphologi G3-ID CHEMICAL IDENTIFICATION PARTICLE SHAPE PARTICLE SIZE PROTEIN

More information

10 REASONS THAT THE ZETASIZER NANO IS CHOSEN FOR SCIENTIFIC SUCCESS

10 REASONS THAT THE ZETASIZER NANO IS CHOSEN FOR SCIENTIFIC SUCCESS 10 REASONS THAT THE ZETASIZER NANO IS CHOSEN FOR SCIENTIFIC SUCCESS Abstract: New components and measurement techniques continue to be introduced for the Zetasizer Nano. New to the system are a Surface

More information

Differentiation of polymer branching and composition using the Mark Houwink plot

Differentiation of polymer branching and composition using the Mark Houwink plot Differentiation of polymer branching and composition using the Mark Houwink plot MOLECULAR SIZE MOLECULAR STRUCTURE MOLECULAR WEIGHT Introduction The manipulation of polymer properties through changes

More information

Overcoming and quantifying Wall Slip in measurements made on a rotational rheometer

Overcoming and quantifying Wall Slip in measurements made on a rotational rheometer Overcoming and quantifying Wall Slip in measurements made on a rotational rheometer RHEOLOGY AND VISCOSITY Introduction When making shear rheological measurements on structured liquids, in particular suspensions,

More information

Nanobubble Applications and Characterization by Nanoparticle Tracking Analysis

Nanobubble Applications and Characterization by Nanoparticle Tracking Analysis Nanobubble Applications and Characterization by Nanoparticle Tracking Analysis Review of ultra-fine bubble (nanobubble) applications and characterization examples PARTICLE CONCENTRATION PARTICLE SIZE Introduction

More information

GPC/SEC Analysis of Polymer Solutions Used in Inkjet Printing

GPC/SEC Analysis of Polymer Solutions Used in Inkjet Printing GPC/SEC Analysis of Polymer Solutions Used in Inkjet Printing Introduction Inkjet printing of polymer solutions is an area of interest for depositing precise, high-throughput patterns of functional polymer.

More information

How to use GPC/SEC for compositional analysis

How to use GPC/SEC for compositional analysis How to use GPC/SEC for compositional analysis Determining the relative concentration of two components in a polymer sample MOLECULAR SIZE MOLECULAR STRUCTURE MOLECULAR WEIGHT Introduction Over the last

More information

Studying the Effect of Steric Layer Thickness on Emulsion Stability

Studying the Effect of Steric Layer Thickness on Emulsion Stability Studying the Effect of Steric Layer Thickness on Emulsion Stability Dr Tim J. Wooster, Professor Mary A. Augustin, Food Science Australia (CSIRO) of Chemistry, Monash University, Victoria 3800, Australia

More information

Determining cure profile and post-cure shrinkage of photopolymers using UV accessory on a rotational rheometer

Determining cure profile and post-cure shrinkage of photopolymers using UV accessory on a rotational rheometer Determining cure profile and post-cure shrinkage of photopolymers using UV accessory on a rotational rheometer RHEOLOGY AND VISCOSITY Introduction A photopolymer usually consists of monomers, oligomers

More information

How switching mobile phases can improve your OMNISEC results

How switching mobile phases can improve your OMNISEC results How switching mobile phases can improve your OMNISEC results Analysis of ester- and acid-capped PLGA samples in mobile phases of DCM and THF MOLECULAR SIZE Introduction MOLECULAR Biodegradable, thermoplastic

More information

Using rheology to study the hardness and spreadability of butter

Using rheology to study the hardness and spreadability of butter Using rheology to study the hardness and spreadability of butter RHEOLOGY AND VISCOSITY Introduction Butter is a multiphase emulsion consisting of fat globules, crystalline fat and an aqueous phase dispersed

More information

Using the Zetasizer Nano to optimize formulation stability

Using the Zetasizer Nano to optimize formulation stability Using the Zetasizer Nano to optimize formulation stability PARTICLE SIZE ZETA POTENTIAL The top five ways you can use the Zetasizer Nano to help improve the stability of your formulation. Formulation development

More information

Evaluating the rheological properties of hyaluronic acid hydrogels for dermal filler applications

Evaluating the rheological properties of hyaluronic acid hydrogels for dermal filler applications Evaluating the rheological properties of hyaluronic acid hydrogels for dermal filler applications RHEOLOGY AND VISCOSITY PARTICLE SIZE Introduction Hyaluronic acid (HA) is a naturally occurring polysaccharide

More information

MALVERN ADVANCED GPC/SEC DETECTORS SETTING THE STANDARD MOLECULAR WEIGHT MOLECULAR SIZE MOLECULAR STRUCTURE

MALVERN ADVANCED GPC/SEC DETECTORS SETTING THE STANDARD MOLECULAR WEIGHT MOLECULAR SIZE MOLECULAR STRUCTURE MOLECULAR WEIGHT MOLECULAR SIZE MOLECULAR STRUCTURE MALVERN ADVANCED GPC/SEC DETECTORS SETTING THE STANDARD Setting the Standard WHAT CAN ADVANCED GPC/SEC DETECTION OFFER YOU? A modern research environment

More information

Measurements of protein electrophoretic mobility using the Zetasizer Nano ZSP

Measurements of protein electrophoretic mobility using the Zetasizer Nano ZSP Measurements of protein electrophoretic mobility using the Zetasizer Nano ZSP Introduction Zetasizer Nano is the market leader in dynamic and electrophoretic light scattering technology for measurements

More information

MEASURING PROTEIN AGGREGATION WITH THE VISCOTEK SEC-MALS 20

MEASURING PROTEIN AGGREGATION WITH THE VISCOTEK SEC-MALS 20 MEASURING PROTEIN AGGREGATION WITH THE VISCOTEK SEC-MALS 20 Introduction Protein aggregation is recognised as a major issue in the biopharmaceutical industry. Proteins have a tendency to aggregate over

More information

The ultimate in desktop particle characterization

The ultimate in desktop particle characterization Particle size Zeta potential Molecular weight ZS The ultimate in desktop particle characterization detailed specification from www.malvern.com/zetasizernano The Zetasizer Nano ZS brings you the practicality

More information

Measuring the rheology of thermoplastic polymer melts

Measuring the rheology of thermoplastic polymer melts Measuring the rheology of thermoplastic polymer melts Using rotational and capillary rheometry to characterize polymer melts RHEOLOGY AND VISCOSITY Introduction Rheology is the science of studying the

More information

Sample measurements to demonstrate Zetasizer specifications (Zetasizer Nano, Zetasizer APS, Zetasizer μv)

Sample measurements to demonstrate Zetasizer specifications (Zetasizer Nano, Zetasizer APS, Zetasizer μv) Sample measurements to demonstrate Zetasizer specifications (Zetasizer Nano, Zetasizer APS, Zetasizer μv) Contents Size using Non-Invasive back-scatter (NIBS) optics Zetasizer Nano S/ZS...2 Minimum size

More information

Zetasizer Nano ZSP: A Perfect Tool For Life Science Applications

Zetasizer Nano ZSP: A Perfect Tool For Life Science Applications Zetasizer Nano ZSP: A Perfect Tool For Life Science Applications Dr Mike Kaszuba Technical Support Manager E-mail: michael.kaszuba@malvern.com Contents Zetasizer Nano ZSP Software Enhancements Protein

More information

The ultimate in desktop particle characterization

The ultimate in desktop particle characterization Particle size Zeta potential Molecular weight ZS The ultimate in desktop particle characterization detailed specification from www.malvern.com/zetasizernano The Zetasizer Nano ZS brings you the practicality

More information

Zeta potential - An introduction in 30 minutes

Zeta potential - An introduction in 30 minutes Zeta potential - An introduction in 30 minutes ZETA POTENTIAL Introduction Zeta potential is a physical property which is exhibited by any particle in suspension, macromolecule or material surface. It

More information

Protein-Protein Interaction Measurement: Balancing Complete Characterization with High- Throughput Capability in Formulation Development

Protein-Protein Interaction Measurement: Balancing Complete Characterization with High- Throughput Capability in Formulation Development Protein-Protein Interaction Measurement: Balancing Complete Characterization with High- Throughput Capability in Formulation Development Introduction High throughput screening in the world of pharmaceutical

More information

ZS90. The ultimate in desktop particle characterization. Particle size. Zeta potential. Molecular weight

ZS90. The ultimate in desktop particle characterization. Particle size. Zeta potential. Molecular weight Particle size Zeta potential Molecular weight ZS90 The ultimate in desktop particle characterization detailed specifications from www.malvern.com/zetasizernano A routine analysis system for particle characterization

More information

Meet Our Uncle: 12 Stability Applications on One Platform

Meet Our Uncle: 12 Stability Applications on One Platform Tech Note Meet Our Uncle: 12 Stability Applications on One Platform Uncle is an all-in-one stability platform that enables twelve different applications with one instrument. Fluorescence, static light

More information

Laser diffraction and automated imaging: complementary techniques for nasal spray development

Laser diffraction and automated imaging: complementary techniques for nasal spray development Laser diffraction and automated imaging: complementary techniques for nasal spray development PARTICLE SIZE PARTICLE SHAPE Together the techniques of laser diffraction and automated particle imaging support

More information

Biophysical characterization of SMALPs and nanodiscs

Biophysical characterization of SMALPs and nanodiscs Biophysical characterization of SMALPs and nanodiscs Verna Frasca, Ph.D. Malvern Panalytical Verna.Frasca@Malvern.com Malvern Panalytical 2 SMALP April 11 2018 Malvern Panalytical Biosciences Group Solutions

More information

The Diffusion Barrier Technique, Practical Aspects and Data interpretation

The Diffusion Barrier Technique, Practical Aspects and Data interpretation The Diffusion Barrier Technique, Practical Aspects and Data interpretation Introduction The diffusion barrier technique is a method for reliably measuring the electrophoretic mobility of proteins by minimising

More information

Meet Stunner: The one-shot protein concentration and sizing combo

Meet Stunner: The one-shot protein concentration and sizing combo TECH NOTE Meet Stunner: The one-shot protein concentration and sizing combo Introduction What if you could get a better read on the quality of your biologics and use less sample at the same time? Stunner

More information

Electrophoretic Light Scattering Overview

Electrophoretic Light Scattering Overview Electrophoretic Light Scattering Overview When an electric field is applied across an electrolytic solution, charged particles suspended in the electrolyte are attracted towards the electrode of opposite

More information

Isothermal experiments characterize time-dependent aggregation and unfolding

Isothermal experiments characterize time-dependent aggregation and unfolding 1 Energy Isothermal experiments characterize time-dependent aggregation and unfolding Technical ote Introduction Kinetic measurements have, for decades, given protein scientists insight into the mechanisms

More information

Measuring the rheology of polymer solutions

Measuring the rheology of polymer solutions Measuring the rheology of polymer solutions John Duffy, Product Marketing Manager, Rheology RHEOLOGY AND VISCOSITY MICRORHEOLOGY Polymers are versatile materials that play an important role in a diverse

More information

ZETASIZER NANO Series PERFORMANCE, SIMPLICITY, VERSATILITY

ZETASIZER NANO Series PERFORMANCE, SIMPLICITY, VERSATILITY ZETASIZER NANO Series PERFORMANCE, SIMPLICITY, VERSATILITY Performance, Simplicity, Versatility ZETASIZER NANO SERIES Research Performance, Operational Simplicity, Application Versatility The Zetasizer

More information

Gold Nanoparticle Applications and Characterization by Nanoparticle Tracking Analysis

Gold Nanoparticle Applications and Characterization by Nanoparticle Tracking Analysis Gold Nanoparticle Applications and Characterization by Nanoparticle Tracking Analysis Review of nanoparticle size and concentration analysis in gold colloid applications PARTICLE SIZE PARTICLE CONCENTRATION

More information

Malvern Macromolecular Solutions. Innovative Solutions in Material Characterization

Malvern Macromolecular Solutions. Innovative Solutions in Material Characterization Malvern Macromolecular Solutions Innovative Solutions in Material Characterization MALVERN HEADQUARTERS Malvern Instruments is a Spectris company. Innovative Solutions in Material Characterization Particle

More information

A basic guide to particle characterization

A basic guide to particle characterization A basic guide to particle characterization Introduction 3 What is a particle? 3 Why measure particle properties? 3 Which particle properties are important to measure? 4 Particle Properties 5 Particle size

More information

KINEXUS DSR SERIES REDEFINING RHEOMETER CAPABILITIES FOR ASPHALT TESTING

KINEXUS DSR SERIES REDEFINING RHEOMETER CAPABILITIES FOR ASPHALT TESTING KINEXUS DSR SERIES REDEFINING RHEOMETER CAPABILITIES FOR ASPHALT TESTING ASPHALT CHARACTERIZATION AND CLASSIFICATION TO ASPHALT INDUSTRY STANDARDS Asphalt, or bitumen, is a petroleum product used in the

More information

next generation Welcome to the Dispersion options Smarter particle sizing Your sample, expertly processed and prepared Particle size

next generation Welcome to the Dispersion options Smarter particle sizing Your sample, expertly processed and prepared Particle size Particle size Welcome to the next generation Your sample, expertly processed and prepared Dispersion options 3000 detailed specification sheets from Getting your sample preparation right The US Pharmacopeia

More information

Generating Aggregation Rates to Determine Stability of Monoclonal Antibodies

Generating Aggregation Rates to Determine Stability of Monoclonal Antibodies Generating Aggregation Rates to Determine Stability of Monoclonal Antibodies Background Static light scattering is widely used to measure the molar masses of molecules, particles and their aggregates under

More information

Inform is a series of white papers designed to provide advice on material characterization issues. Mie theory The rst 100 years

Inform is a series of white papers designed to provide advice on material characterization issues. Mie theory The rst 100 years Inform is a series of white papers designed to provide advice on material characterization issues Mie theory The rst 100 years Mie theory the rst 100 years One hundred years ago Gustav Mie developed an

More information

CONFOCHECK. Innovation with Integrity. Infrared Protein Analysis FT-IR

CONFOCHECK. Innovation with Integrity. Infrared Protein Analysis FT-IR CONFOCHECK Infrared Protein Analysis Innovation with Integrity FT-IR CONFOCHECK: FT-IR System for Protein Analytics FT-IR Protein Analysis Infrared spectroscopy measures molecular vibrations due to the

More information

A Basic Guide to INFORM WHITE PAPER. Abstract:

A Basic Guide to INFORM WHITE PAPER. Abstract: A Basic Guide to Abstract: The aim of this guide is to provide you with a basic grounding in the main particle characterization techniques currently in use within industry and academia. It assumes no prior

More information

Litesizer Sheet 1. [Home]

Litesizer Sheet 1. [Home] Litesizer 500 15.01.15 Sheet 1 [Home] The Product: Litesizer 500 Measurement Modes Particle Size & Distribution Dynamic light scattering (DLS) Zeta Potential Electrophoretic light scattering (ELS) Molecular

More information

Investigating the Relationship Between the Rheological Properties of Hyaluronic Acid and its Molecular Weight and Structure using Multidetector

Investigating the Relationship Between the Rheological Properties of Hyaluronic Acid and its Molecular Weight and Structure using Multidetector Investigating the Relationship Between the Rheological Properties of Hyaluronic Acid and its Molecular Weight and Structure using Multidetector SEC and SEC-MALS Presented by Bassem Sabagh, PhD Technical

More information

Bohlin. Rheological instruments backed with rheological experience. Rheological properties

Bohlin. Rheological instruments backed with rheological experience. Rheological properties Rheological properties ṙ Bohlin Rheological instruments backed with rheological experience detailed specification sheets from /bohlingemini The Bohlin Gemini rheometers are an advanced range of compact

More information

Particle Analysis at the Touch of a Button. Litesizer series

Particle Analysis at the Touch of a Button. Litesizer series Particle Analysis at the Touch of a Button Litesizer series Particle systems can be complex The size and stability of nanoparticles and microparticles are crucial to their function, as well as to their

More information

Highly automated protein formulation development: a case study

Highly automated protein formulation development: a case study Application Note Highly automated protein formulation development: a case study Introduction and background Therapeutic proteins can be inherently prone to degradation and instability, and they often pose

More information

Introduction to Dynamic Light Scattering with Applications. Onofrio Annunziata Department of Chemistry Texas Christian University Fort Worth, TX, USA

Introduction to Dynamic Light Scattering with Applications. Onofrio Annunziata Department of Chemistry Texas Christian University Fort Worth, TX, USA Introduction to Dynamic Light Scattering with Applications Onofrio Annunziata Department of Chemistry Texas Christian University Fort Worth, TX, USA Outline Introduction to dynamic light scattering Particle

More information

REVERSE ENGINEERING PHARMACEUTICAL FORMULATION USING THE MORPHOLOGI G3-ID.

REVERSE ENGINEERING PHARMACEUTICAL FORMULATION USING THE MORPHOLOGI G3-ID. REVERSE ENGINEERING PHARMACEUTICAL FORMULATION USING THE MORPHOLOGI G3-ID. Introduction According to the definition established by the FDA, a generic drug is a drug product which is comparable to a reference

More information

Morphologically directed chemical identification Coupling particle characterization by morphological imaging with Raman spectroscopy

Morphologically directed chemical identification Coupling particle characterization by morphological imaging with Raman spectroscopy Inform is a series of white papers designed to provide advice on material characterization issues Morphologically directed chemical identification Coupling particle characterization by morphological imaging

More information

One-stop stability. T m & T agg. T m with SYPRO (DSF) Isothermal stability. Thermal recovery. Sizing & polydispersity. Sizing with thermal ramp B 22

One-stop stability. T m & T agg. T m with SYPRO (DSF) Isothermal stability. Thermal recovery. Sizing & polydispersity. Sizing with thermal ramp B 22 One-stop stability Cracking stability using a pile of one-trick, protein-hungry tools is a ton of work. UNcle combines 3 different measurement modes fluorescence, SLS and DLS. So you can crank out all

More information

KINEXUS SERIES REDEFINING RHEOMETER CAPABILITIES

KINEXUS SERIES REDEFINING RHEOMETER CAPABILITIES KINEXUS SERIES REDEFINING RHEOMETER CAPABILITIES KINEXUS SERIES Redefining rheometer capabilities for characterizing dispersed systems At Malvern, we haven t just redesigned a rheometer we ve redefined

More information

KINEXUS SERIES REDEFINING RHEOMETER CAPABILITIES

KINEXUS SERIES REDEFINING RHEOMETER CAPABILITIES KINEXUS SERIES REDEFINING RHEOMETER CAPABILITIES KINEXUS SERIES Redefining rheometer capabilities for characterizing dispersed systems At Malvern, we haven t just redesigned a rheometer we ve redefined

More information

Artisan Technology Group is your source for quality new and certified-used/pre-owned equipment

Artisan Technology Group is your source for quality new and certified-used/pre-owned equipment Artisan Technology Group is your source for quality new and certified-used/pre-owned equipment FAST SHIPPING AND DELIVERY TENS OF THOUSANDS OF IN-STOCK ITEMS EQUIPMENT DEMOS HUNDREDS OF MANUFACTURERS SUPPORTED

More information

Page 1 of 5. Is it alright to estimate dñ/dc in SLS measurements?

Page 1 of 5. Is it alright to estimate dñ/dc in SLS measurements? Page 1 of 5 Is it alright to estimate dñ/dc in SLS measurements? Due to the complexity of measuring the specific refractive index increment (dñ/dc), static light scattering molecular weight measurements

More information

Introduction to Dynamic Light Scattering for Particle Size Determination

Introduction to Dynamic Light Scattering for Particle Size Determination www.horiba.com/us/particle Jeffrey Bodycomb, Ph.D. Introduction to Dynamic Light Scattering for Particle Size Determination 2016 HORIBA, Ltd. All rights reserved. 1 Sizing Techniques 0.001 0.01 0.1 1 10

More information

YOUR PARTNER IN PARTICLE CHARACTERIZATION

YOUR PARTNER IN PARTICLE CHARACTERIZATION YOUR PARTNER IN PARTICLE CHARACTERIZATION SERVICES escubed limited provides comprehensive analytical and research and development services in colloidal, particulate and powder technology to customers across

More information

Measuring Lysozyme Monomer at 0.1 mg/ml Concentration. Equipment used : Sample Preparation and Measurement :

Measuring Lysozyme Monomer at 0.1 mg/ml Concentration. Equipment used : Sample Preparation and Measurement : Application Report #001 Measuring Lysozyme Monomer at 0.1 mg/ml Concentration Equipment used : ALV-NIBS / HPPS High Sensitivity Version, Lysozyme (MERCK), 0.1 molar Sodium-Acetate buffer (ph 4.25), syringe

More information

Suspension Stability; Why Particle Size, Zeta Potential and Rheology are Important

Suspension Stability; Why Particle Size, Zeta Potential and Rheology are Important ANNUAL TRANSACTIONS OF THE NORDIC RHEOLOGY SOCIETY, VOL. 20, 2012 Suspension Stability; Why Particle Size, Zeta Potential and Rheology are Important Mats Larsson 1, Adrian Hill 2, and John Duffy 2 1 Malvern

More information

Chirascan 6-Cell Peltier Cell Holder: Rapid Optimisation of Buffer Conditions for Stabilising Protein Therapeutics

Chirascan 6-Cell Peltier Cell Holder: Rapid Optimisation of Buffer Conditions for Stabilising Protein Therapeutics CHIRASCAN SERIES APPLICATION NOTE Chirascan 6-Cell Peltier Cell Holder: Rapid Optimisation of Buffer Conditions for Stabilising Protein Therapeutics Abstract: The selection of buffer conditions that maximise

More information

Novel instrumentation to gain insight into the aggregation state of proteins by Taylor Dispersion Analysis: a comparison

Novel instrumentation to gain insight into the aggregation state of proteins by Taylor Dispersion Analysis: a comparison Novel instrumentation to gain insight into the aggregation state of proteins by Taylor Dispersion Analysis: a comparison with Dynamic Light Scattering Wendy Louise Hulse *, Rob Forbes, David Goodall, Alex

More information

South pacific Journal of Technology and Science

South pacific Journal of Technology and Science Zetasizer Technique Dr. Nagham Mahmood Aljamali Abstract: Assist. Professor in Organic Chemistry, Chemistry Department.,College of Education.,Kufa University.,IRAQ. In this review study., zetasizer technique

More information

ORTHOGONAL PARTICLE CHARACTERIZATION TECHNIQUES FOR BIO-APPLICATIONS: AN INTRODUCTION TO DLS (DYNAMIC LIGHT SCATTERING), NTA (NANOPARTICLE

ORTHOGONAL PARTICLE CHARACTERIZATION TECHNIQUES FOR BIO-APPLICATIONS: AN INTRODUCTION TO DLS (DYNAMIC LIGHT SCATTERING), NTA (NANOPARTICLE ORTHOGONAL PARTICLE CHARACTERIZATION TECHNIQUES FOR BIO-APPLICATIONS: AN INTRODUCTION TO DLS (DYNAMIC LIGHT SCATTERING), NTA (NANOPARTICLE TRACKING ANALYSIS) AND RMM (RESONANT MASS MEASUREMENT) - Dr. Markus

More information

Analytical methods to characterize aggregates and subvisible particles

Analytical methods to characterize aggregates and subvisible particles Analytical methods to characterize aggregates and subvisible particles Andrea Hawe 9 th P4EU Meeting 30 th November 2015 * MPI * Martinsried 1 Background of company privately held, independent service

More information

LS Spectrometer. The DLS and SLS system for sophisticated research

LS Spectrometer. The DLS and SLS system for sophisticated research LS Spectrometer The DLS and SLS system for sophisticated research P r o f e s s i o n a l L i g h t S c a t t e r i n g S o l u t i o n s LS INSTRUMENTS THE GLOBAL LEADER IN ADVANCED LIGHT SCATTERING TECHNOLOGIES

More information

Particle Analysis at the Touch of a Button. Litesizer series

Particle Analysis at the Touch of a Button. Litesizer series Particle Analysis at the Touch of a Button Litesizer series Particle systems can be complex The size and stability of nanoparticles and microparticles are crucial to their function, as well as to their

More information

The Illumination of Structure using Light Scattering. Michael Caves Product Technical Specialist for Biophysical Characterisation

The Illumination of Structure using Light Scattering. Michael Caves Product Technical Specialist for Biophysical Characterisation The Illumination of Structure using Light Scattering Michael Caves Product Technical Specialist for Biophysical Characterisation Light Scattering Laser Scattered light Detector Scattered light intensity

More information

Setting the Standard for Advanced GPC/SEC Systems

Setting the Standard for Advanced GPC/SEC Systems Absolute molecular weight Molecular conformation Molecular size Solution viscosity Viscotek Setting the Standard for s ṙ ṙ Detailed specification sheets from Introducing the system - the leading solution

More information

Holographic Characterization of Protein Aggregates

Holographic Characterization of Protein Aggregates Holographic Characterization of Protein Aggregates Size, morphology and differentiation one particle at a time (and fast) D. G. Grier, C. Wang, X. Zhong, & M. D. Ward New York University D. B. Ruffner

More information

Positive and Nondestructive Identification of Acrylic-Based Coatings

Positive and Nondestructive Identification of Acrylic-Based Coatings Positive and Nondestructive Identification of Acrylic-Based Coatings Using Partial Least Squares Discriminant Analysis with the Agilent 4300 Handheld FTIR Application Note Materials Testing and Research

More information

Using runt Disposable Cuvettes

Using runt Disposable Cuvettes Tech Note Guide Table of Contents Ease of Use... 3 Sensitivity... 4 Resolution... 5 Accuracy... 6 Faster Sample Prep... 7 Measuring Molecular Weight... 8 Observing Protein Aggregation... 9 Characterize

More information

Analytical Ultracentrifugation Macromolecule Characterization in Solution

Analytical Ultracentrifugation Macromolecule Characterization in Solution Analytical Ultracentrifugation Macromolecule Characterization in Solution AUC TECH OVERVIEW OPTIMA AUC Analytical Ultracentrifugation Beckman Coulter delivered the first AUC sample characterization tool

More information

Advantages of Agilent AdvanceBio SEC Columns for Biopharmaceutical Analysis

Advantages of Agilent AdvanceBio SEC Columns for Biopharmaceutical Analysis Advantages of Agilent AdvanceBio SEC Columns for Biopharmaceutical Analysis Comparing Columns from Different Vendors to Improve Data Quality Technical Overview Introduction Size exclusion chromatography

More information

Lateral Flow: Making Magnetic Particles A Viable And Easier To Use Replacement To Gold Nanoparticles.

Lateral Flow: Making Magnetic Particles A Viable And Easier To Use Replacement To Gold Nanoparticles. AN2 LATERAL FLOW COUPLING KIT Lateral Flow: Making Magnetic Particles A Viable And Easier To Use Replacement To Gold Nanoparticles. Introduction While lateral flow immunoassays represent a quick, low cost,

More information

Analysis of demicellization data from isothermal titration calorimetry

Analysis of demicellization data from isothermal titration calorimetry Analysis of demicellization data from isothermal titration calorimetry MICROCALORIMETRY Introduction Many industrial and biochemical applications depend on the usage of detergents and other surfactants.

More information

Monitoring the Aggregation of Particles through Raman Spectroscopy

Monitoring the Aggregation of Particles through Raman Spectroscopy Monitoring the Aggregation of Particles through Raman Spectroscopy Name: Yanxiao Ma Advisor: Dr. Andrew Callender Submission Date: 19 Oct. 2012 Introduction Raman spectroscopy is a spectroscopic technique

More information

Surface Zeta Potential Cell (ZEN1020)

Surface Zeta Potential Cell (ZEN1020) Surface Zeta Potential Cell (ZEN1020) MAN0483 Issue 1.0 September 2011 English Copyright Malvern Instruments Ltd. 2011 Malvern Instruments makes every effort to ensure that this document is correct. However,

More information

Dr. Christoph Johann Wyatt Technology Europe GmbH Copyright Wyatt Technology Europe GmbH All Rights reserved 1

Dr. Christoph Johann Wyatt Technology Europe GmbH Copyright Wyatt Technology Europe GmbH All Rights reserved 1 Dr. Christoph Johann Wyatt Technology Europe GmbH 2010 Copyright Wyatt Technology Europe GmbH All Rights reserved 1 Introduction Overview The Nature of Scattered Light: Intensity of scattered light Angular

More information

Particle Characterization Laboratories, Inc.

Particle Characterization Laboratories, Inc. Analytical services Particle size analysis Dynamic Light Scattering Static Light Scattering Sedimentation Diffraction Zeta Potential Analysis Single Point Titration Isoelectric point determination Aqueous

More information

Use of SEC-MALS. (Size Exclusion Chromatography - Multi Angle. Light Scattering) for protein quality and characterization

Use of SEC-MALS. (Size Exclusion Chromatography - Multi Angle. Light Scattering) for protein quality and characterization Use of SEC-MALS (Size Exclusion Chromatography - Multi Angle Light Scattering) for protein quality and characterization Methods for protein characterization Analytical SEC is a common method to characterize

More information

Amended Specifications as per the Pre Bid meeting held on in Chapter 4

Amended Specifications as per the Pre Bid meeting held on in Chapter 4 NIPER-G/PUR/GLOBAL/2018/06 Tender Specification for Nano Particle Size (DLS based), Zeta Potential, Molecular weight, Micro rheology measurement A proven complete system should have the measurement facility

More information

Clemens Stilling Characterisation, Analytical Sciences for Biologicals, UCB Pharma S.A. Alun, living with Parkinson s disease

Clemens Stilling Characterisation, Analytical Sciences for Biologicals, UCB Pharma S.A. Alun, living with Parkinson s disease Clemens Stilling Characterisation, Analytical Sciences for Biologicals, UCB Pharma S.A. Alun, living with Parkinson s disease Outline Short intro Aggregate characterisation during candidate selection and

More information

Holographic Characterization of Agglomerates in CMP Slurries

Holographic Characterization of Agglomerates in CMP Slurries Holographic Characterization of Agglomerates in CMP Slurries Total Holographic Characterization (THC) Comparison of THC to other technologies Dynamic Light Scattering (DLS) Scanning Electron Microscopy

More information

Electronic Supplementary Material (ESI) for Nanoscale This journal is The Royal Society of Chemistry Characterization of CeO2 NP suspensions

Electronic Supplementary Material (ESI) for Nanoscale This journal is The Royal Society of Chemistry Characterization of CeO2 NP suspensions % Vol. Electronic Supplementary Material (ESI) for Nanoscale Characterization of CeO2 NP suspensions CeO2 NPs show a strong tendency to aggregate as a function of ph, ionic strength, dilution and sonication

More information

Precision and accuracy of protein size determination using the ActiPix TDA200 Nano-Sizing System

Precision and accuracy of protein size determination using the ActiPix TDA200 Nano-Sizing System Precision and accuracy of protein size determination using the ActiPix TDA200 Nano-Sizing System Keywords: Hydrodynamic radius, diffusion coefficient, sizing, Taylor dispersion analysis, protein, antibodies,

More information

Gel Permeation Chromatography (GPC) or Size Exclusion Chromatography (SEC)

Gel Permeation Chromatography (GPC) or Size Exclusion Chromatography (SEC) Gel Permeation Chromatography (GPC) or Size Exclusion Chromatography (SEC) Size Exclusion Chromatography (SEC) is a non-interaction based separation mechanism in which compounds are retained for different

More information

Static and dynamic light scattering. Cy Jeffries EMBL Hamburg

Static and dynamic light scattering. Cy Jeffries EMBL Hamburg Static and dynamic light scattering. Cy Jeffries EMBL Hamburg Introduction. The electromagnetic spectrum. visible 10-16 10-10 10-8 10-4 10-2 10 4 (l m) g-rays X-rays UV IR micro wave Long radio waves 400

More information

Accessory Publication

Accessory Publication 10.1071/CH10199_AC CSIRO 2010 Australian Journal of Chemistry, 2010, 63(9), 1326 1329 Accessory Publication Self-Assembly of Protoporphyrin IX-TEG Derivatives into Tunable Nanoscaled Spherical Structures

More information

Impurities In Biomolecules Institute for International Research. Monitoring & Predicting Biomolecular Aggregation Using Light Scattering

Impurities In Biomolecules Institute for International Research. Monitoring & Predicting Biomolecular Aggregation Using Light Scattering Impurities In Biomolecules Institute for International Research Monitoring & Predicting Biomolecular Aggregation Using Light Scattering Kevin Mattison Malvern Instruments Ewa Folta-Stogniew Yale University

More information