Application note December 15th, 2022

HCP analysis: ELISA or Mass Spectrometry

Host cell proteins (HCPs) are inside a cell and used for recombinant protein expression. HCPs can contaminate biopharmaceutical products, with the effect of reducing the effectiveness of the drug or causing patients to have non-wanted reactions. That is why their detection is crucial. The quantification of HCPs can be done using both ELISA and Mass Spectrometry. What is the difference between these methods? Which one should be preferred?



What is the ELISA test?

ELISA test principle

ELISA is short for enzyme-linked immunosorbent assay and is one of the first tests used in medical diagnostics. ELISA test can quickly determine the presence or absence of specific biological molecules, such as antibodies or antigens, from just a few samples. ELISA test works by tagging a molecule and then immersing it into skin tissue or blood. If there are same-type molecules present in the sample, they will bind to the tagged molecule and be measured to determine their levels. ELISA testing provides scientists with valuable insight into numerous scientific breakthroughs, from uncovering new treatments for illnesses to developing new ways to detect cancer cells earlier than ever before. ELISA test is an invaluable tool when it comes to healthcare research.


HCP analysis with ELISA

ELISA tests are specifically used in the analysis of Host Cell Proteins (HCP) in biopharmaceutical products to ensure safety and efficacy. ELISA tests can detect the presence of HCPs at very low concentrations by using antigen-antibody interactions between the ELISA reactive components and the actual HCPs. The ELISA test can detect variations of protein integrity related to manufacturing process interference, making it a reliable tool when it comes to analysing protein samples. ELISA testing has become a powerful method that researchers resort to over more traditional methods due to its versatility, high throughputs and relatively simple experimental setup.

ELISA is often used for HCP detection tests as it can provide high: 

  • throughput;
  • sensitivity;
  • selectivity.


What is Mass Spectrometry?

Principle of Mass Spectrometry (MS)

Mass spectrometry is an instrumental technique that allows scientists to analyze the mass and chemical structure of a given sample. This technique works by ionizing a sample, breaking the molecules into smaller parts so each mass can be identified. These mass spectra are generated by accelerating particles through an electric and/or magnetic field, allowing scientists to obtain mass-to-charge ratios which can then be used to figure out the mass of atoms or molecules within the sample. MS also provides detailed information on how much of each particular mass is present within a sample, making it perfect for use in drug testing and other forensic research. Thanks to mass spectrometry, researchers have more tools than ever to understand their samples – and uncover secrets once thought completely unreachable!

Mass Spectrometry for the detection of HCPs

Mass Spectrometry plays a major role when it comes to analyzing host cell proteins (HCPs). This analytical technique is especially valuable, as it can quickly detect impurities in pharmaceutical biologic products. Mass spectrometry works by ionizing the compounds and then separating them based on their mass-to-charge ratio, enabling researchers to identify HCPs at a molecular level with great accuracy. Through this analysis, scientists can determine certain characteristics of the sample such as whether or not there is an increase/decrease in the concentration of particular proteins over time. Mass spectrometry also can distinguish various types of HCPs, and allows identification and quantification of all detected HCPs, which allows for more detailed analysis and interpretation of complex protein samples.

ELISA versus Mass Spectrometry: Comparison

Host Cell Protein analysis is a critical step for biopharmaceutical companies to ensure the safety and quality of their product. ELISA and Mass Spectrometry are two of the most widely used HCP detection methods and have each been proven to be reliable in providing accurate results.

 ELISA is known for its convenience, allowing large samples to be tested in a short period. However, it tends to provide less consistent results than Mass Spectrometry when dealing with complex samples. 

On the other hand, Mass Spectrometry has the advantage of being able to differentiate between HCPs from different species, granting higher levels of specificity. It measures a wider range of parameters than ELISA. The main downside is that it takes longer to process samples due to their complexity and precision requirements.

Finally, these two methods are complementary. ELISA is used to quantify major HCPs without identification of what is really quantified, but with a high throughput and ease of use. MS, thanks to HCP profiler solution, is able to identify and quantify  HCPs individually which allows a full characterisation of all HCPs at each step of the bioprocess.


Besides, this technique is now highly recommended by the authorities.

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