Separation Sciences and Analytical Biotechnology Laboratory

Separation sciences and analytical biotechnology

At the Centre for Vaccine Evaluation (CVE), our expertise in separation sciences and analytical biotechnology helps provide new tools for characterizing and analysing biologics.

 

Why we study biologics using separation sciences and analytical biotechnology

Biologics such as vaccines, blood products, and recombinant therapeutic protein preparations can contain simple or complex substances. As the amount, combination, and characteristics of these substances affect how the biotherapeutic product will act in the body, evaluating the quality of biologics is important. We use separation sciences and analytical biotechnology to analyse biologics for quality control. We also use them to identify and characterize the substances present and to detect contaminants. Furthermore, we use separation sciences and analytical biotechnology to investigate whether a formulation matches its description and whether active ingredients meet quality standards.

As we develop and improve analytical techniques, we acquire tools for evaluating biologics. New tools help us analyse complex mixtures and structures, detect contaminants, and examine new biomolecules and biologics.

 

How we study biologics using separation sciences and analytical biotechnology

At the Centre for Vaccine Evaluation (CVE), in the Separation Sciences and Analytical Biotechnology Laboratory, we develop and improve methods for separating and analysing substances in biologics. Analysis can be challenging because of the complexity of mixtures and structures. We need to separate small amounts of active ingredients, such as proteins, from samples that contain large amounts of added ingredients (excipients) such as diluents and carrier ingredients.

We use technically advanced instruments and separation methods to investigate and apply high-resolution analytical techniques. For example, we use capillary electrophoresis and high-performance liquid chromatography (HPLC) to study biomolecules and address concerns such as product quality, purity, contaminant detection, and batch consistency.

We apply our expertise and analytical abilities to:

  • Develop and improve techniques in order to analyse sample characteristics such as product quality, purity, and batch consistency;
  • Generate research information that will help establish criteria and standards for quality and purity testing of biologics as well as lot-release testing programs;
  • Develop and improve techniques to study subsequent entry biologics;
  • Analyse the quality of biologics to provide information, such as impurity profiles and contaminant detection, for assessing and responding to adverse reactions;
  • Provide a Canadian perspective for setting international standards and methods for biological substances by participating in international collaborative studies with other regulatory agencies such as the U.S. Food and Drug Administration, World Health Organization, European Pharmacopoeia, and United States Pharmacopeia;
  • Provide support to manufacturers for setting up and optimizing high-resolution separation techniques.

 

Concepts and tools we use in the separation sciences and analytical biotechnology laboratory

Capillary electrophoresis is a powerful approach that we use at the Centre for Vaccine Evaluation (CVE) to separate and characterize charged molecules, large or small, such as DNA and proteins. Samples are run through an electric field in a thin tube, of less than 100 micrometers, called capillary tubing. In the capillary electrophoresis instrument, light sources acting as detectors track and measure how molecules migrate or move.

Since capillary electrophoresis uses small capillary tubing, it has advantages over standard electrophoresis methods, such as:

  • Enabling the use of high voltages without damaging samples;
  • Providing faster processing by using increased voltages, sensitivity or detection;
  • Using light detection measurements;
  • Analysing small amounts of sample;
  • Automating and analysing large sets of samples relatively quickly.

High-performance liquid chromatography (HPLC) is a powerful tool that we use at CVE to separate, identify and measure the amount of a substance in a sample. In general, the sample is prepared by dissolving it in an appropriate solvent and running it through a separation column under high pressure. Compounds from the sample move through the column at different speeds as a result of their affinity to bind to the column material. Different compounds have different affinities and separate from each other. When a compound leaves the column, it goes through a UV light and a UV detector measures the UV light absorbed by the sample. Then, we use the collected and processed data to characterize the compounds in the sample. In addition, we can couple HPLC with mass spectrometry to more completely characterize the sample.

 

Research highlight 1: Development of new approaches to characterizing erythropoietin samples

Erythropoietin (EPO) is a glycoprotein hormone that controls the production of red blood cells in the human body. In humans, it is produced by the kidneys and liver. Since the mid-1980s, manufacturers have been able to generate EPO using recombinant DNA technology. This recombinant human erythropoietin (rhEPO) provided a lifeline to millions of people around the world, who were suffering from anemia or other conditions following adverse events from chemotherapy or AIDS-related therapies.

Therapeutic formulations of erythropoietin contain large amounts of non-therapeutic ingredients (they are usually called excipients) that stabilize the active ingredient. Human serum albumin (HSA) is a protein frequently used in pharmaceutical products as an excipient. In some instances, products contain 30 times as much HSA as rhEPO. Such large amounts of extra components make it challenging to analyse and characterize erythropoietin formulations.

In our laboratory, we address this concern by investigating the use of capillary electrophoresis to analyse various erythropoietin formulations and batches. We have developed an effective way to resolve and characterize erythropoietin in the untreated formulation. As we do not need to treat the sample before analysis, we do not need to remove, derive or modify components.

We are working to:

  • Develop new methods and approaches to examining final erythropoietin products;
  • Detect and characterize quality and quantity of erythropoietin in samples;
  • Develop new tools for surveillance of marketed products;
  • Develop new tools for distinguishing closely related erythropoietin;
  • Develop new tools for assessing subsequent entry biologics as new erythropoietin preparations are developed and proposed as therapeutic products.

 

Research highlight 2: Characterization of somatropin (Human Growth Hormone) - setting new standards and tools for evaluating subsequent entry biologics and marketed products

Somatropin, also known as human growth hormone, is a protein produced by the pituitary gland of the human body that plays a major role in growth regulation. Recent advances in technology, such as recombinant DNA technology, have enabled the production of biosynthetic human growth hormone for therapeutic applications.

As biologics, somatropin preparations are analysed for quality using a wide range of specific standard tests to characterize the substances present in the sample to maximize their safety. Then, the biologics are compared against a standard to confirm specific biological activity based on protein structure and formulation.

In our laboratory, we use modern technology to characterize the structure of somatropin and develop better ways to analyse new and marketed products. We have used high-performance liquid chromatography (HPLC) combined with other analytical approaches to develop an effective approach to examine somatropin samples. In the course of this work, we have identified impurities that had been unreported previously.

We are working to:

  • Identify and characterize somatropin;
  • Detect and characterize contaminants such as somatropin degradation forms and structural variants;
  • Develop new methods and approaches to examine somatropin samples;
  • Develop new tools for surveillance of marketed products;
  • Develop new tools to assess subsequent entry biologics as new somatropins are developed and proposed as therapeutic products.

 

For information about the lead scientist of this laboratory, please visit their  Directory of Scientists and Professionals profile.

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