1. CO-OP REFLECTION PRESENTATION
PROTEIN ENGINEERING AND ANTIBODY TECHNOLOGIES-
ANTIBODY DISCOVERY GROUP
SAMRITI BEDI
SUPERVISOR- JUKKA KONOLA
RESEARCH ASSISTANT CO-OP- JULY- DECEMBER 2014
EMD SERONO, BILLERICA

2. Outline
Overall work flow at Antibody Technology, EMD Serono
Primary role and responsibilities as a Co-op student
Assay development and optimization on 2 major formats:
ELISA
HTRF FRET
Accomplishments
Acknowledgements

3. 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

4. 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.

5. 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

6. Automated ELISA system

7. 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 Tag
Target Protein
Antibody coupled with
cryptate (Donor) Antibody coupled with
D2(Acceptor)
Laser/Flash Lamp excitation
at 320nm
Detect the energy transfer
by reading at 665nm

8. Accomplishments
Learned 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.

9. 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

10. Thank you!