SmartEnzymes at BGI, an interview with Aaron Bailey

We got the opportunity to talk to Aaron Bailey at BGI Americas. From their newly setup MS labs, they use SmartEnzymes to help provide analytical services to their customers globally. Read on to find out more.

 
 Tell us about yourself and BGI?

Currently I work at BGI Americas as the Associate Director of Product Management for Mass Spectrometry Services. I am part of the scientific team based in San Jose, California, at the newly built Mass Spectrometry Center.

 
 How has the MS team set up the industrial platform there at BGI in San Jose?

Our mass spec center is a service and R&D laboratory focused on providing LC-MS solutions for proteomic research and biologic drug characterization. Our current LC-MS service platform plays to our core strengths in automated sample prep, multi-mode HPLC separations, high resolution Orbitrap MS, and advanced bioinformatic analysis. We have built a versatile system which can cater to a wide scope of proteomics and biologic drug characterization needs. In San Jose our Q Exactive HF-X BioPharma can be coupled to either nano-flow or analytical flow UHPLC, which essentially affords us the ability to provide LC-MS services for multi-omic researchers and pharmaceutical research and development.

 
 How does the MS team do sample preparation prior to MS analysis?

For emerging formats, including some Fc-fusion protein drugs which are already established in global markets, glycosylation can be both highly heterogeneous and often involve both N- and O-linkages and is often distributed on potentially many sites throughout the amino acid sequence. These projects require that glycosylations can be selectively removed in order to validate the identity of each identified PTM. We have had success using several deglycosidases including SmartEnzymes from Genovis. In a sort of combinatorial sample prep matrix these enzymes help objectively determine for each therapeutic protein how highly complex glycoform patterns obseved at the (completely unprocessed) intact mass level are specifically derived from the contributions of possibly several types of glycosylation events (i.e., N-linked vs. O-linked vs. sialylated). Each may individually involve heterogeneous distibution of the specific sugar types found at any particular amino acid site. While protein isoform distributions must be measured at the “intact” or “subunit” level, for example using FabRICATOR, site specific measurements must be performed using peptide mapping approaches. Most importantly, the two types of datasets must essentially agree on the findings, and the sample prep strategy described here is a powerful way to navigate this challenge.

 
 What are the advantages of using SmartEnzymes like FragIT, OglyZOR and SialEXO?

We found that the activity and purity of these SmartEnzymes was a good fit for our native LC-MS platform in the sense that we are able to efficiently deglycosylate our therapeutic proteins using very little amount enzyme which remains near or below the actual level of detection in our experiments. This means that in normal usage we are not seeing these enzymes (FragIT, OglyZOR and SialEXO) in our datasets, which is particularly important for our native LC-MS intact mass analyses. This is a critical point for us as it means that we can keep our analyses streamlined by avoiding the need to address any artifact data which are not specifically related to any therapeutic protein isoforms.

The San Jose Mass Spectrometry Center uniquely allows BGI to offer state-of-the-art genomic and proteomic analytical services globally. In the coming year we plan to grow our team and add new LC and MS instrumentation to further increase our ability to support this market. We expect to see exciting new developments in integrating NGS and LC-MS datasets. We also plan to introduce additional mass spectrometry services which will allow us to provide even deeper support in our main focus areas of proteomics and biologics characterization.

 
 Download Aaron’s ASMS poster here.

 
 For more information on FragIT, OglyZOR and SialEXO please visit the following pages:

• FragIT, OglyZOR and SialEXO Product Pages
• Generation of Antibody Fragments & Analyis of O-glycans