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Screening and Subcloning of High Producer Transfectomas Using Semisolid Media and Automated Colony Picker

  • Suba Dharshanan
  • Cheah Swee Hung
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1131)

Abstract

Generation of high-producing clones is a perquisite for achieving recombinant protein yields suitable for biopharmaceutical production. However, in many industrially important cell lines used to produce recombinant proteins such as Chinese hamster ovary, mouse myeloma line (NS0), and hybridomas, only a minority of clones show significantly above-average productivity. Thus, in order to have a reasonable probability of finding rare high-producing clones, a large number of clones need to be screened. Limiting dilution cloning is the most commonly used method, owing to its relative simplicity and low cost. However the use of liquid media in this method makes the selection of monoclonal hybridoma and transfectoma colonies to be labor intensive and time consuming, thus significantly limiting the number of clones that can be feasibly screened. Hence, we describe the use of semisolid media to immobilize clones and a high-throughput, automated colony picker (ClonePix FL) to efficiently isolate monoclonal high-producing clones secreting monoclonal antibodies.

Key words

High-throughput selection Mammalian cells Monoclonal antibodies Recombinant proteins Semisolid media 

Notes

Acknowledgment

This work was supported by Inno Biologics and Ministry of Science, Technology and Innovation, Malaysia.

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Suba Dharshanan
    • 1
  • Cheah Swee Hung
    • 2
  1. 1.Protein Science DepartmentInno BiologicsNilaiMalaysia
  2. 2.Faculty of Medicine, Department of PhysiologyUniversity of MalayaKuala LumpurMalaysia

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