FEB 11, 2021 10:00 AM PST

Intact and peptide level characterization of nanobodies by high resolution HPLC-MS/MS

Sponsored by: SCIEX
Speaker
  • Head of Technology at EpiQMAx
    Biography
      Peng Yu received his Bsc in Environmental Engineering from Zhejiang University and PhD in plant biology from the University of Minnesota. He subsequently conducted postdoctoral research in chemical proteomics as a Humboldt fellow with Prof. Bernhard Küster at the Technical University of Munich.
      He is currently responsible for growing and overseeing the biologics+ service branch at EpiQMAx GmbH. He has also served as the head of infectious disease mass spec lab at Novartis and tech lead of glycoproteomics biomarker discovery platform at InterVenn Biosciences in the past.

    Abstract
    Date:  February 11, 2021
    Time: 10:00am (PST),  1:00pm (EST)
     
    Nanobodies are single-domain antibodies derived from heavy-chain-only antibodies (HcAbs) commonly found in camelids, consisting solely of the variable, antigen-binding domain. Their small size affords some unique properties such as higher stability, better tissue penetration with potential access to cryptic tumors and easier, higher yield prokaryotic production, which has attracted growing interest in therapeutic and research use.
     
    In this work, we characterized the stochastic labeling of two research-grade nanobodies using the benchtop X500B QTOF LC-MS/MS System. The former nanobody contains an Alexa Fluor dye conjugated via the maleimide linker, while the latter nanobody is biotin-labeled via N-hydroxysuccinimide (NHS) conjugation. A quick and reproducible intact mass measurement was able to confirm the ID, purity, modifications and disulfide bonds. The differences in heterogeneity in labeling between maleimide and NHS chemistry is also readily seen. A rapid trypsin digestion followed by peptide mapping was carried out to pinpoint the specific sites and degrees of labeling. Finally, we were able to adjust the biotin labeling efficiency by varying reagent to nanobody ratio . The expression, purification and characterization of these research-grade nanobodies lay the groundwork for future therapeutic programs.
     
    Learning Objectives
    • Nanobodies as an alternative to monoclonal IgGs and its unique properties and advantages
    • Facile characterization of nanobodies and their functional derivatives using LC-MS
    • Fine tuning of product properties guided by LC-MS data
     
     
    Webinars will be available for unlimited on-demand viewing after live event.

    Show Resources
    You May Also Like
    DEC 02, 2020 8:00 AM PST
    C.E. CREDITS
    DEC 02, 2020 8:00 AM PST
    DATE: December 2nd, 2020 TIME: 08:00am PDT, 11:00pm EDT Bioreactors and shakers are used to cultivate microorganisms, plant, insect, and mammalian cells in different volumes. Upscaling of pr...
    OCT 08, 2020 7:00 AM PDT
    C.E. CREDITS
    OCT 08, 2020 7:00 AM PDT
    DATE: October 8, 2020 TIME: 7:00am PDT, 10:00am EDT, 4:00pm CEST How often do you pipette in your cell culture lab every day? Usually, we do it so often that we tend stop thinking about ho...
    NOV 18, 2020 8:00 AM PST
    C.E. CREDITS
    NOV 18, 2020 8:00 AM PST
    DATE: November 18, 2020 TIME: 08:00am PDT We develop and implement technologies to solve some of the major bottlenecks in biomedical research. In particular, we establish new imaging approac...
    NOV 16, 2020 8:00 AM PST
    C.E. CREDITS
    NOV 16, 2020 8:00 AM PST
    Date: November 16, 2020 Time: 8:00am (PST), 11:00am (EST) CRISPR screening has become the prime discovery tool in modern biomedical research and drug discovery. At the same time, most screen...
    OCT 29, 2020 6:00 AM PDT
    C.E. CREDITS
    OCT 29, 2020 6:00 AM PDT
    Date: October 29, 2020 Time: 6:00am (PDT), 9:00am (EDT), Chronic inflammation can occur as a result of a combination of genetic predispositions and environmental factors. Epigenetic modifica...
    SEP 10, 2020 9:00 AM PDT
    C.E. CREDITS
    SEP 10, 2020 9:00 AM PDT
    Date: September 10, 2020 Time: 9:00am (PDT), 12:00pm (EDT) Osmolality testing is relevant throughout the entire bioprocessing workflow. As customers look to refine mAb and gene therapy workf...
    Loading Comments...
    Show Resources
    Attendees
    • See more