Figure 1: Measuring particle mass in a fluid Figure 2: Configuration of a Resonance Mass Spectrometer (Malvern Archimedes System) Figure 3: (from left to right) (a), (b), and (c): Aggregate detection using the Malvern Archimedes system Figure 4: Demonstration of the ability to distinguish between protein aggregates and contaminated silicone oil droplets Multifunctional waterproof makeup bag, waterproof shopping bag, waterproof wash bag, waterproof mommy bag Dongguan Jinying Handbags Co.,Ltd , https://www.jinyingbag.com
Application of Resonance Quality Test Method in Biopharmaceutical Development
**Contributor:** Malvern Instruments
**Title:** Application of the Resonance Mass Measurement Method in Biopharmaceutical Development
**Author:** Dr. Lisa Newey-Keane, Biopharmaceutical Product Manager at Malvern Instruments
**Company Number:** MAL/JOB/2720
**Word Count:** 2286
**Image:** 4 images in total (available at the end of the file; high-resolution files available separately)
**Figure 1:** Measuring particle mass in a fluid
**Figure 2:** Configuration of the resonance mass test instrument (Archimedes system, Malvern Instruments)
**Figure 3:** (from left to right) (a), (b), and (c): Aggregate detection using the Malvern Archimedes system
**Figure 4:** Demonstration of the ability to distinguish between protein aggregates and contaminated silicone oil droplets
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**Application of the Resonance Mass Measurement Method in Biopharmaceutical Development**
**Author:** Dr. Lisa Newey-Keane, Biopharmaceutical Product Manager at Malvern Instruments
In the rapidly evolving field of biopharmaceuticals, predicting and measuring protein aggregation remains one of the most significant challenges. Dr. Lisa Newey-Keane from Malvern Instruments has introduced a novel analytical method that offers deeper insights into the behavior of protein aggregates during formulation development.
With the increasing investment in biomolecular research, the demand for more accurate and reliable analytical techniques has never been higher. These developments are not only expensive but also tightly regulated, which makes it essential to have robust methods for quality control and data collection. Many industry professionals highlight that analytical bottlenecks are a growing concern, potentially hindering drug development if not addressed effectively.
Unlike small molecule drugs, biopharmaceuticals are complex, non-synthetic, and often non-crystalline, making their purity and efficacy more difficult to assess. Impurities such as aggregates, misfolded proteins, or denatured molecules can significantly impact product performance. This complexity demands specialized analytical tools that differ from those used in traditional pharmaceutical development, presenting new challenges for both manufacturers and regulators.
The selection of candidate molecules involves various physicochemical tests to identify potential issues early on. One critical question is how these molecules will behave once formulated. Protein aggregates are particularly concerning due to their potential to trigger immune responses, making their detection and quantification essential.
Regulatory agencies have expressed concerns about protein aggregates ranging from 0.2 to 2 microns in size, yet current particle sizing techniques lack the sensitivity to provide quantitative data in this range. The Resonance Mass Measurement (RMM) technique, implemented through the Malvern Archimedes system, addresses this gap by enabling the detection and counting of particles from 50 nm to 5 microns in diameter.
**Technical Overview**
Resonance Mass Measurement relies on a mechanical resonator that detects changes in mass. When particles pass through the microfluidic channel of the sensor, they alter the resonant frequency of the structure. This change is measured with high precision, allowing for accurate mass determination.
For example, when particles enter the resonator, the frequency shifts, reaching a maximum at the end of the channel before returning to baseline as the particles exit. By measuring this frequency shift, the mass of individual particles can be calculated.
This method is especially useful for detecting buoyant particles like oil droplets, which cause a positive frequency shift compared to the baseline. The Malvern Archimedes system uses MEMS-based sensors with microfluidic channels to detect even the smallest particles. Each sensor measures the frequency changes caused by the flow of the sample, providing detailed information about particle mass, size, and concentration.
**Quantitative Analysis of Protein Aggregates**
Initially, protein aggregates form at the dimer level, eventually growing to sizes up to tens of microns. Traditional methods like flow microscopy are limited in their ability to analyze sub-micron particles. RMM fills this gap by offering precise quantification of particles in this size range.
Figure 3(a) shows the particle size distribution of submicron IgG aggregates in a 4 μL buffer solution. Using RMM, the concentration of aggregates larger than 300 nm was found to be 4 x 10ⶠper mL. The mass-based measurements allow for the calculation of aggregate composition in terms of the number of protein molecules involved.
After applying shear stress, Figure 3(c) demonstrates an increase in aggregate concentration from 300 nm to 1 μm, indicating a tenfold rise. This suggests an aggregate cascade, which is often associated with poor formulation stability under stress.
**Pollutant Detection: Silicone Oil Droplets**
Silicone oil droplets are commonly found in formulations due to their use as lubricants in syringes and containers. These droplets are similar in size to protein aggregates, making them difficult to distinguish using conventional methods. However, RMM can differentiate between the two based on buoyancy.
As shown in Figure 4, protein aggregates—being denser than the buffer—produce negative peaks in the frequency trajectory. In contrast, silicone oil droplets, being less dense, create positive peaks. This distinction ensures more accurate results and avoids false positives.
**Current Applications and Future Potential**
One of the key advantages of RMM is its ability to work with minimal sample volume—just 100 μL of a 100 cP viscous sample—without requiring extensive preparation. This makes it ideal for analyzing valuable biological materials.
Due to its accuracy in detecting and quantifying particles across a wide size range, RMM is becoming a preferred tool in early-stage biologics development. It supports the concept of "early discovery, lower cost," helping companies identify issues before they become costly problems.
In summary, the Resonance Mass Measurement method represents a powerful advancement in biopharmaceutical analysis, offering greater precision, reliability, and insight into the behavior of protein aggregates and contaminants.
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