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

Automated ELISA system

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

Thank you!