What is Hybrid LC-MS/MS?

Technically Hybrid LC-MS/MS is a technique that combines an affinity capture step with LC-MS/MS detection. Usually it only requires one antibody, whereas a typical LBA technique will require two. Following affinity capture, it is common to include a digestion step to produce surrogate peptides, which are representative of the protein of interest and are used for quantitation. The overall process takes advantage of the combined selectivity of the extraction and the flexibility of tandem mass spectrometry to differentiate the analyte from other potential interferents.

How long has KCAS been utilizing LC-MS/MS?

KCAS initiated the use of hybrid LC-MS/MS for the quantitation of large molecules over three years ago. We have experts in this field with over 20-years’ experience who launched this service to bridge the gap that exists in supporting large molecule modalities, and we employ several different technologies to do this. We have considerable experience with several modalities; from therapeutic proteins to fusion proteins, to antibodies, biomarkers and ADCs.

How can Hybrid LC-MS/MS benefit you?

Hybrid LC-MS/MS can benefit you and your project’s needs because it can represent the best choice for many scenarios. Depending on availability of reagents it might be the only analytical option. Often, we want to be able to differentiate what is sometimes a very small sequence difference between similar moieties. In a situation where there are only one or two amino acid differences, the digestion can be optimized to select peptides that contain the sequential difference so the molecular masses and fragmentation will be easily differentiated using LC-MS/MS. We have worked on other projects where no reagents were available making LC-MS/MS most likely the only bioanalytical option. In a recent example, we were developing an assay for an ADC where there was no antibody to the payload. In this example we used LC-MS/MS to detect the peptide containing the payload for a quick and efficient approach where we could not have developed an assay at all using LBA.

Why is KCAS the best choice for your LC-MS/MS project?

At KCAS, we work with our clients to understand the specific project drivers and what is needed to support their drug development requirements. We try to evaluate each project by starting with feasibility of LC-MS/MS, LBA, or both. We can do this in series, or in parallel, depending on the urgency of the project. We like to be able to evaluate these complimentary technologies so we can “pivot” with the client if needed based on the quality of data that each approach provides. Since we are all in the same building, sharing the same labs, this process is made much easier than it can be with an organization using a more siloed approach across multiple locations.

We believe by having significant expertise with both technologies, it eliminates process bias and allows us to find the best answer for our clients. It is possible that both technologies can work, but sometimes one is clearly the better choice. The specialists at KCAS can help you ensure you know all the options before conducting your project.

We look forward to evaluating your project with you soon! Please use the form below to let us know when you are available to meet!