samriti bedi co-op reflection presentation- northeastern university
TRANSCRIPT
CO-OP REFLECTION PRESENTATION
PROTEIN ENGINEERING AND ANTIBODY TECHNOLOGIES-ANTIBODY DISCOVERY GROUP
S A M R I T I B E D IS U P E RV I S O R - J U K K A KO N O L AR E S E A R C H A S S I S TA N T C O - O P- J U LY- D EC E M B E R 2 0 1 4E M D S E R O N O, B I L L E R I C A
OutlineOverall work flow at Antibody Technology, EMD SeronoPrimary role and responsibilities as a Co-op studentAssay development and optimization on 2 major formats:ELISAHTRF FRET
AccomplishmentsAcknowledgements
Overall Workflow
3
Antigen Immunization
B cell sorting and Culturing/Cloning
Possible IgG/IgM generation
Capture ELISA to test for presence/absence of IgG/IgMs
Target Binding ELISA to identify target specific IgGs
Identification of clones producing target specific IgGs
Confirmation and further characterization
Protein production
Primary role and responsibilities
Developing and performing validation assays for testing reagents including target proteins and antibodies by binding ELISA.
Preparing 384/96 well plates coated with target proteins for a binding ELISA.
Screening of samples by a fully automated target binding ELISA to identify target specific IgGs.
Screening of samples by a semi-automatic or fully automatic capture ELISA to test for IgG/IgM production by B cells.
Performing serum titer experiments on samples by fully automatic target binding ELISA.
MVS Performance Verification Testing of the Perkin Elmer Sciclone Liquid Handler systems.
Developing and optimizing HTRF FRET assays for different antibody-target systems.
Maintaining Sciclone automated liquid handler systems by Perkin Elmer on a day-to-day basis.
Maintaining an up-to-date record of the electronic scientific notebook.
Maintaining a database of reagent supplies within the automation lab.
ELISA Assays Application:
Target binding
Ig expression determination
Serum titer determination
Isotype determination
Reagent validation assays
Target
Primary antibody against target protein
Antibody against Fc part of the primary Ab coupled with HRP
TMB Addition
TMB turns into a blue product – and stop reagent turns it yellow for reading at 450nm
HTRF Technology Homogeneous Time Resolved Fluorescence• Immunoassay based on transfer of energy between 2 fluorophores• a donor e.g. Eu3+ or Lumi4®-Tb cryptate• an acceptor e.g. d2
• The donor/acceptor used in HTRF emit long-lived fluorescence and hence the short lived background signals decay
6-His TagTarget Protein
Antibody coupled with cryptate (Donor) Antibody coupled with
D2(Acceptor)
Laser/Flash Lamp excitation at 320nm
Energy transfer to donor when in close proximity
Detect the energy transfer by reading at 665nm
AccomplishmentsLearned to work in a corporate environment under strict EHS guidelines.
Significantly helped to move company projects further by continuously delivering results in a timely and accurate manner.
Developed an acumen for multi-tasking and time management to meet project deadlines.
Worked independently and also in-collaboration with other group members to run various high-throughput assays.
Developed a good understanding of the basics of the immunoassays used.
Gained experience at analyzing and presenting scientific data succinctly.
Acknowledgements
Antibody Technology Group at EMD Serono: Jukka Konola Youbin Wang Xinyan Zhao
Xiubin Gu Qingyong Ji
An Qi Jinyang Zhang Shruti Pratapa Christel Iffland
Northeastern University: Vanecia Harrison Cynthia Bainton
Jim Leung Other Co-op Students:
Miriam-El-Samin Sneha Ranade