Researchers at the New Jersey Institute of Technology (NJIT) have unveiled a new lab technique that they believe represents a “paradigm shift” in the way pharmaceutical labs test and manufacture new protein-based drugs, such as therapeutic monoclonal antibodies being developed to to treat a variety of diseases, from cancer to infectious diseases.
Researchers say their electrochemistry-based approach, described in the journal Analytical Chemistrycould enable safety and quality testing of emerging biotherapeutics to be performed in a fraction of the time required for conventional methods, which typically require the lengthy and expensive production of certain biomaterials used for sample testing.
The study was conducted in collaboration with researchers from Merck, Johnson & Johnson and Ohio University.
“This method that we developed at NJIT has the potential to have a major impact in quantitative proteomics, and it represents a paradigm shift in the pharmaceutical industry in terms of monitoring biopharma product and process impurities for quality control,” says Hao Chen, the paper’s corresponding author and professor in the Department of Chemistry and Environmental Sciences at NJIT.
“With this study, we have now demonstrated an approach that can quantify and process drug impurities much faster and more accurately than had been possible… We expect it to become very useful in facilitating the development of therapeutic proteins and vaccines for the treatment and prevention of various diseases in the future.”
Traditionally, such testing, or protein quantification, has involved the time-consuming preparation of synthetic isotope-tagged peptides that are used as internal standards to measure total protein concentrations in a sample, allowing researchers to actively monitor the efficacy and safety of therapeutic protein components during the development of proteins. the drug method.
To overcome this limitation, Chen’s lab developed a coulometric mass spectrometry (CMS) approach for absolute quantification of proteins without the use of standards. The method instead applies liquid chromatography-mass spectrometry and an electrochemical flow cell to rapidly quantify and detect changes in target proteins or peptides based on electrochemical signatures.
“Instead of waiting weeks to obtain standards or reagents in traditional approaches, one could perform CMS quantification experiments right away. Thus, it would facilitate tracking of impurities of drugs discovered during the process and ensure their effective removal with process optimization and control,” Chen said.
“Such a device allows us to separate peptides after protein digestion with liquid chromatography, monitor peptide oxidation in the electrochemical flow cell to produce an electric current, and measure the oxidation yield by mass spectrometry,” explained the first author. of the article and NJIT Ph.D. . student Yongling Ai. “The combination of electrical current signals along with the oxidation yield provides sufficient information for absolute quantification of peptides and proteins.”
In their study, the team demonstrated its CMS method by achieving absolute quantification of multiple proteins (β-lactoglobulin B, -lactalbumin and carbonic anhydrase) in a mixture in a single run, without using standards.
In particular, the team also showed the method’s capabilities for detecting protein deamidation – a common degradation event in therapeutic proteins due to physical or chemical stress during manufacturing and storage.
The team successfully quantified several protein degradation products, including an important intermediate of protein degradation — the formation of succinimide — which, according to the study authors, has never been done before with absolute quantification due to a lack of standards.
“The lack of standards is caused by the challenges in their de novo synthesis,” Chen said. “Being able to accurately quantify the deamidation products and intermediates could provide a better understanding of therapeutic protein degradation and potentially create a new way to investigate disease pathologies and aging processes.”
Now Chen’s lab plans to apply their new method for large-scale quantification of thousands of proteins in one run. They also plan to improve the sensitivity of their CMS analysis to quantify very low levels of proteins in complex biological samples, which could benefit research efforts ranging from clinical diagnostics and drug discovery to precision medicine for which identification and quantification of samples at the molecular level is necessary.
“Because proteins perform a wide variety of functions within organisms, the importance of absolute protein quantification is difficult to overestimate,” Chen said. “CMS should accelerate processes for disease diagnosis, discovery and drug development, and it now opens a new door for biologists and biochemists to investigate amounts of proteins in the human body that may fulfill important biological functions or roles as biomarkers for disease and targets for medicines. ”
Researchers develop a simpler and faster way to quantify and explore therapeutic proteins
Yongling Ai et al, Standard free absolute quantification of antibody deamidation degradation and host cell proteins by coulometric mass spectrometry, Analytical Chemistry (2022). DOI: 10.1021/acs.analchem.2c02709
Quote: Researchers Unlock a New Method for Testing Protein-Based Drugs (2022, Sept. 27) Retrieved Sept. 27, 2022 from https://phys.org/news/2022-09-method-protein-based-drugs.html
This document is copyrighted. Other than fair dealing for personal study or research, nothing may be reproduced without written permission. The content is provided for informational purposes only.