Process Related Impurity Analysis for Biotherapeutics

Impurity testingGyrolab® systems and immunoassays speed up the bioprocessing workflow

Process related impurity concentrations are highly regulated in the manufacture of biologic drugs because of their immunogenic potential. Bioanalytical methods to detect and measure process related impurities must be developed carefully to characterize heterogeneous mixtures of host cell proteins (HCPs) present in samples or the carryover of culture or process impurities such as Protein A along with the therapeutic.

  • Heterogeneous mixtures of host cell proteins (HCPs), for example
    CHO-HCP, E. coli HCP, PLBL-2 and transferrin
  • Process-specific impurities
  • Carryover of culture or process impurities such as Protein A and insulin

Impurity analysis using current ELISA technologies have limitations of labor-intensive protocols, lower throughput and narrow dynamic range. As a result, repeat analysis is common, further delaying project decisions and lot approvals. Automating impurity analysis with Gyrolab miniaturized, microfluidic immunoassays provides fast turn-around time, broad dynamic ranges and reproducibility, and high-quality data, with less hands-on operations offering an ideal technology to support all phases of bioprocess and production for biotherapeutics.

Gyrolab immunoassays provide faster time to data compared to traditional ELISA methods

Gyrolab immunoassays provide faster time to data compared to traditional ELISA methods (data courtesy of Medimmune, the global biologics research and development arm of AstraZeneca).

HCP analysis with expanded dynamic range

One primary goal during HCP assay development is a wide dynamic range for optimized analyte detection, avoiding repeat analysis and additional sample dilutions. A wide dynamic range is especially important for bioprocess HCP analysis, since the concentration range of host cell protein impurities encountered in process samples may be especially wide. Gyrolab assays and kits offer broad analytical ranges, reducing the the number of dilutions required to hit the analytical range, thereby reducing the need for repeat runs. Gyrolab CHO-HCP E3G Kit, (broad dynamic range of ~3 ng/mL – 8000 ng/mL), and Gyrolab CHO-HCP Kit 1 both incorporate antibodies from Cygnus Technologies.

Gyrolab CHO HCP Kit 1

Gyrolab CHO-HCP Kit 1 offers shows a broad analytical range of approximately 2 – 8000 ng/mL compared to 1 – 1000 ng/mL for ELISA.

In situations where assay development with process-specific reagents is required, Gyrolab Bioaffy™ 1000 HC Assay Toolbox provides the flexibility, automation and rapid time to results.

Protein A and culture impurities

Other bioprocess impurities may carry over from purification columns or culture media additives. Gyrolab Protein A kits detect GE Healthcare MabSelect SuRe™ ligands in samples, and carefully tested Gyrolab assay protocols are available for Human Transferrin, Insulin, and Phospholipase B-Like 2 (PLBL2) impurity analysis. The wider analytical range of Gyrolab immunoassays offers several logs working range expansion over ELISA for a wider detection range requiring a lower MRD.

protein A

Ready-to-use Gyrolab Protein A kit covers a broader dynamic range compared to a commercial ELISA kit.

Comparable to ELISA

Gyrolab assays have been shown by many of our customers to be comparable to ELISAs. For example, the correlation of process samples during purification of a therapeutic protein analyzed by both ELISA and Gyrolab immunoassays is shown below.

Correlation-of-Gyrolab-with-ELISA-s--1Correlation of results for samples throughput the purification of a therapeutic protein are show. The error bars indicate that the result from Gyrolab xP workstation are less variable.

Gyrolab immunoassay platform for process impurity analysis

table impurity-1


Host-Cell Protein Analysis to Support Downstream Process DevelopmentLearn more about how scientists at Sanofi have successfully implemented Gyrolab immunoassay system as a reliable platform for high-throughput HCP analysis, in combination with a robotic liquid handling system. The results were comparable to their gold-standard ELISA method.

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    Gyrolab Systems
    Gyrolab technology reduces manual labor, assay development time, time to result and material consumption -and it improves workflow efficiency. 

    Gyrolab Kits, CDs and Rexxip Buffers
    Gyrolab Kits, CDs and Rexxip Buffers - Designed for best results 

    Gyrolab Protein A Kit
    Gyrolab Protein A Kit Quantifies MabSelect SuRe™ ligands and native Protein A residuals in bioprocess samples. 
    [Product information sheet]

    Gyrolab CHO-HCP Kit 1
    Gyrolab CHO-HCP Kit 1 is designed to measure CHO Host Cell Proteins impurities with performance that addresses many of the limitations of ELISA. 
    [Product information sheet]

    Quantitation of Host Cell Protein Impurities
    Gyrolab HCP assays deliver high precision results over a broad dynamic range, minimizing repeat analysis in bioprocess development and production. 
    [Application Note]

    Comparative HCP Testing Using Traditional and Emerging Technologies
    Martha Jackson, David Cirelli, Phoebe Baldus, Ned Mozier. BioTherapeutics Pharmaceutical Sciences, Analytical Research and Development, Pfizer, Andover, MA 

    High Throughput Process-related Impurity Testing Using a Gyrolab™ Workstation
    Meletios Roussis, Eric Beard, Keshavamurthy Prakash, Robert Strouse and Michael Washabaugh. Analytical Biochemistry, MedImmune, Gaithersburg, MD 20878 USA 

    Development of residual host cell protein assays for recombinant microbial biopharmaceuticals
    Katja Peiris and Daniel Schiwek, LCMD Analytical Development, Lonza AG, Switzerland 

    Minimizing Repeats and Making Rapid Data Driven Decisions using Automated Nanoliter-scale Immunoassays for Impurity Testing in Bioprocess and Production
    This recorded webinar focuses on the use of automated, nanoliter-scale flow-through technology to address a critical need for improved speed, throughput and reduced repeats in process related impurity testing. Presenters: Martha L. Jackson, Senior Research Scientist, Pfizer Inc. and Jun Hyuk Heo, Biochemist, Biotechnology New and Enabling Technologies (BNET), Merck & Co. 

    Cobra Biologics on the versatility of Gyrolab™ immunoassay platform in bioprocess development
    Anna Lönnerbro is an Analytical Scientist at Cobra Biologics, a CMO and CRO of biologics and pharmaceuticals for pre-clinical, clinical and commercial supply. Cobra Biologics offers support from small scale-up to large-scale bioprocess. Presenting at Gyrolab User Seminar in Copenhagen, Denmark on June 2, 2014, Anna described how the Gyrolab platform enables Cobra Biologics to deliver premium services with improved immunoassays.  
    [Case Study]

    How to get more reliable results with less effort
    Analytical results that you can depend on are crucial to success, especially in a regulated environment. For example, immunoassays that consistently deliver reliable data with high accuracy and precision are worth their weight in gold. ELISA has been the standard for immunoassays, but this situation is changing. We present here two case studies that show how a move from ELISA to Gyrolab™ assays is helping your colleagues to improve their results and lab workflows. 
    [Case Study]

    Jun Heo on using Gyrolab xP to automate high throughput HCP analysis
    Recorded Seminar: Jun Heo, Analytical Biochemist, Biologics-New and Enabling Technologies (BNET), Merck & Co., Inc., presents 'The Automation of a High Throughput Residual Assay to Support In-Process Analysis of Monoclonal Antibodies'. 

    Daniel Schiwek on The value of Gyrolab xP workstation in HCP analysis and bioprocess
    Recorded Seminar: Daniel Schiwek, Project Evaluation Manager at Global Biological Operations, Lonza AG, Visp, Switzerland, presents ‘Biopharmaceutical manufacturing in microbial systems – Gyrolab as an analytical platform supporting process development'. 

    Automated nanoliter-scale immunoassays to maximize productivity in bioprocess development
    Recorded Webinar: Karolina Österlund, Manager Field Application Scientists EU and Asia, Gyros Protein Technologies AB, presents Gyrolab™ systems and a range of easy-to-use immunoassay kits as an automated, high-throughput platform for cell line and bioprocess development, illustrated by case studies from Shire, MedImmune and Merck. 

    Streamlined and automated immunoassays adapted for bioprocess workflows accelerate QbD strategy initiatives
    Recorded Webinar: Presenter Daniel Forsström, application scientist at Gyros Protein Technologies  

    Gyrolab CHO-HCP E3G Kit
    For detection of CHO Host Cell Proteins using the industry standard Cygnus CHO HCP 3G ELISA antibody. 
    [Product information sheet]

    Gyrolab E. coli HCP Kit
    For the detection of Host Cell Proteins from Escherichia coli expression systems 
    [Product information sheet]

    Gyrolab Bioaffy 1000 HC Assay Toolbox
    Please download the product information sheet Gyrolab Bioaffy 1000 HC Assay Toolbox 
    [Product information sheet]

    Host-Cell Protein Analysis to Support Downstream Process Development
    Sanofi scientists present the successful implementation of the Gyrolab immunoassay system as a reliable platform for high-throughput HCP analysis, in combination with a robotic liquid handling system. The results were comparable to their gold-standard ELISA method. 
    [Case Study]

    Overcoming Limitations of ELISAs in Process-related Impurity Testing
    In this webinar, Utku Hasbay at Takeda Pharmaceuticals presented transfer of an HCP ELISA to Gyrolab® to expand the dynamic range with a 2 hour assay time 

    Accelerating bioprocess workflows with microfluidic immunoassay-based parallel impurity analysis
    Maximize performance and productivity in your bioprocess workflows for biologics Gyrolab® immunoassays deliver rapid turnaround to support data-driven decision-making 

    High Throughput Platform and Automated Sample Preparation for Host Cell Protein Analysis to Support Downstream Process Development
    Speaker: Suzana Petrovic Analytical Scientist Sanofi. Learn how a high-throughput immunoassay platform supported accelerated bioprocess HCP assay development utilizing Gyrolab xP and CHO-HCP E3G kit.