Ligand binding assays and LC-MS/MS systems are common bioanalytical techniques used for analyzing drug products. LC-MS methods are HPLC assays with MS detection units. Advances in detection systems have resulted in the combination of HPLC systems with two MS detector units. Similarly, over the years ligand binding assays have continuously evolved to provide excellent sensitivity for the analysis of therapeutic compounds in biological samples. However, an increase in complexities and demands gave rise to the need for a more efficient bioanalytical tool. Recently, hybrid LC-MS/MS systems have demonstrated promising results for biomarkers and biotherapeutics.
So let us dive deep into the working of hybrid LC-MS/MS systems.
What is hybrid LC-MS/MS?
As the name suggests, hybrid LC-MS/MS is a combination of traditional ligand binding assays (LBA) and LC-MS/MS analysis. Combining both these techniques gives the edge of selective isolation of biotherapeutic of LBA assays and effective detection and quantitation properties of LC-MS/MS assays. Hybrid LC-MS/MS also needs conventional sample preparation or affinity capture enrichment approaches. However, certain cases can be performed directly through a direct digestion approach.
With the ever-increasing complexities concerning drug development processes such as antibody-drug conjugates, fusion proteins, and pegylated proteins, the biopharmaceutical industry constantly faces an uphill task. Here, the combination of ligand binding assay and LC-MS/MS analysis can help unearth crucial PK/PD properties early in the drug development process. LC-MS/MS systems can simultaneously characterize multiple analytes in a given biological matrix. This added benefit gives a window of opportunity to assess the metabolism and catabolism characteristics of both active and inactive components of the construct and can facilitate additional information for LBA analysis. Let us now explore some crucial advantageous hybrid LC-MS/MS systems can have over conventional ELISA assays.
Primary advantages of Hybrid LC-MS/MS
In general, hybrid LC-MS/MS assays have several benefits over their traditional counterparts. Some primary benefits of hybrid LC-MS/MS assays are as follows:
- Enhanced selectivity and structural data
- Intrinsic physicochemical property-based quantitative results
- Does not need specialized reagents in method development
- Facilitates analysis of multiple analytes in a given study sample
- Needs minimum dilutions and have a wide dynamic assay range
- Fewer matrix interferences
- Less sensitive to cross-reactivity
- The presence of LC-MS/MS system allows the separation of chemically and structurally similar analytes
- The prevailing use of LC-MS/MS in bioanalytical labs further facilitates the shift to hybrid LC-MS/MS systems
Must Read: Good Clinical Practices in the Bioanalytical Laboratory
The road ahead for Hybrid LC-MS/MS systems
There are several challenges around the development of small and large molecule drug products. Out of these challenges, selectivity and specificity are the most critical aspects of drug development. Due to being easy to use and high-throughput analysis, ligand binding assays have a special place in clinical and biological research. However, increasing efficacy and safety demands have resulted in the use of more sensitive assays such as LC-MS/MS analysis. Hence, the hybrid between LBA and LC-MS/MS systems is a promising solution for major drug development challenges. Despite these advantages, similar to HPLC method validation and LC-MS method validation, hybrid LC-MS/MS analysis needs thorough validation for its routine use in drug discovery and development.
