Our portfolio of instruments to aid in biopharmaceutical research has grown significantly in recent years, thanks to innovation from our Bioscience Development Initiative as well as acquisitions of NanoSight, Viscotek and most recently, MicroCal. As a result,the amount of useful resources we have created for those interested in analyzing and characterizing proteins  has also exploded.

This list guides you through 20 essential resources for the protein scientist at all stages of research & formulation development. I hope you find it useful. To explore our full, free archive of articles, whitepapers, application notes and recorded webinars, check out our resource center.

Articles

Protein aggregation1) ‘Critical to Quality’ measurements of protein aggregation in biopharmaceuticals – Developing stable formulations of proteins is crucial to successful drug delivery, efficacy and lack of immunogenicity. Size Exclusion Chromatography (SEC), Dynamic Light Scattering (DLS) and Resonant Mass Measurement (RMM) techniques provide valuable insight into a proteins behaviour in formulation.

2) Characterization of sub-nanometre particle interactions – Nottingham Trent University have used the Zetasizer Nano to demonstrate the predominantly electrostatic nature of protein-aluminium interactions, giving the possibility of building novel aluminium-composite materials using biological processes.

3) Tools for evaluating the stability of human recombinant albumins used in human therapeutics – Aggregation and misfolding during storage is a major challenge in the development of proteins for therapeutic applications. SEC is excellent for detailed characterization, and DLS provides a rapid measurement of many samples in a short time.

Whitepapers

protein crystallography4) Developing a bioformulation stability profile – Preformulation screening of candidate biotherapeutic molecules for stability using a combination of sub-micron light scattering analysis and microcapillary viscometry enables important development decisions to be made earlier in the pipeline.

5) Application of Dynamic Light Scattering (DLS) to Protein Therapeutic Formulations: Principles, Measurements and Analysis (4 part series)

a) Basic Principles – An overview of the key principles of Dynamic Light Scattering: theory, correlation statistics, deconvolution algorithms, and the intensity to mass transform.

b) Concentration Effects and Particle Interactions –  The influence of concentration effects and particle interactions on DLS results, with a roadmap for identifying and distinguishing each type of concentration effect.

c) DLS Deconvolution Algorithms – The basic types of DLS deconvolution algorithms used to extract the intensity weighted particle size distribution from the measured correlogram.

d) FAQs –  Frequently asked questions related to the application of DLS to the characterization of protein therapeutic formulations

Application Notes

Pegylated Protein6) Differentiation and characterization of subvisible particulates in therapeutic protein products – This application note provides an example of how Archimedes can be used to detect and quantify the formation of protein sub-visible particles and the introduction of silicone oil droplets, in response to shear stress.

7) Accelerating the development of optimized therapeutic protein formulations using Differential Scanning Calorimetry – DSC data is useful in predicting the stability of a protein in solution. The Tm indicates thermostability, and Tm determination in different formulations is an approximate measure of the susceptibility to aggregation and other irreversible changes at lower temperatures.

8) PEGylated proteins: Optimizing conjugation reactions using triple detection SEC -This work was performed by Celares GMBH, Germany with samples provided by DIGNA BIOTECH, Spain. In this application note, interferon α-5 was conjugated with two different PEG molecules in different reactions. The individual protein and PEG molecules, as well as the PEGylated conjugates are characterized by SEC and their composition, molecular weight and intrinsic viscosity measured.

9) Overcoming the negative effect of protein structure on molecular weight measurement by Size Exclusion Chromatography – Two protein case studies with Anthrolysin and Bcl-2 -Work performed in conjunction with the Institut Pasteur, Paris, France. A description of how advanced GPC/SEC detection can overcome inaccuracy in the measurement of molecular weight caused by deviation in the structure of the sample protein from that of the protein standards or interaction with the chromatography column matrix.

10) Direct visualization, sizing and counting of protein aggregates using NTA – In this application note, we look at how the Nanoparticle Tracking Analysis (NTA) has been used to measure particle size and concentration in order to monitor protein aggregation over time and in response to thermal stress.

11) Sizing proteins and small molecules in the presence of large particles – This application note evaluates protein and small molecule sizing analysis by the Viscosizer 200 and explains how this technology complements light scattering technologies in the protein characterization workflow.

Webinars

DLS from protein molecules12) Protein aggregation – Bridging the sub micron gap – This webinar provides an overview of the current techniques available for studying subvisible particles, and demonstrates the unique abilities of the Archimedes system in characterizing and quantifying particles in a size range that has, up to now, been a challenge to the biopharmaceutical industry.

13) Power in numbers – Learn more about your protein’s behavior and stability in formulation by adding light scattering functionality to your SEC system – When developing a
formulation stability profile for a biotherapeutics, one of the primary techniques used to identify the oligomeric state and presence or absence of aggregates is size exclusion chromatography (SEC). Traditionally, this technique provides a quantitative analysis of a sample by relating the retention volume of peaks present in the chromatogram to a calibration curve providing a basic identification of the relative molecular weight of each of the peaks.

14) A new approach to improving early characterization of therapeutic formulations – The accurate size measurement of both large and small therapeutic molecules within formulations has long been considered a challenge, complicated by the presence of excipient molecules and high background “noise”. This webinar demonstrates how Malvern’s Viscosizer 200 can see through these complications to measure only the molecules of interest, both accurately and repeatedly.

15) Combining dynamic light scattering and raman spectroscopy to achieve new insights into protein stability, aggregation and high order structure –  This presentation describes a new analytical tool that can work with small amounts of protein samples to rapidly and simultaneously measure relative protein molecular structure and size (including aggregation) over a range of concentrations and formulation conditions. The technique uniquely combines two well-established analytical techniques namely dynamic light scattering (DLS) and Raman spectroscopy to derive structural, thermodynamic and kinetic insights into the mechanisms of protein aggregation and the factors that influence protein stability.

Videos

16) Principles of isothermal titration calorimetry

17) OMNISEC for advanced protein characterization

18) Protein mobility measurements on the Zetasizer Nano ZSP

19) Automated low volume size and viscosity analysis – Viscosizer 200

20) Nanoparticle Tracking Analysis – NanoSight in 90 Seconds