Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2681:291-311.
doi: 10.1007/978-1-0716-3279-6_16.

Bulk Reformatting of Antibody Fragments Displayed on the Surface of Yeast Cells to Final IgG Format for Mammalian Production

Stefania C Carrara  1   2 , Jan P Bogen  1   2 , David Fiebig  1   2 , Julius Grzeschik  3 , Björn Hock  1   4 , Harald Kolmar  5   6
Affiliations

Bulk Reformatting of Antibody Fragments Displayed on the Surface of Yeast Cells to Final IgG Format for Mammalian Production

Stefania C Carrara et al. Methods Mol Biol. 2023.

Abstract

While yeast surface display (YSD) has gained traction for antibody hit discovery efforts with the first therapeutic YSD-isolated antibody sintilimab approved in 2018, a major drawback that remains is the time-consuming reformatting of monoclonal antibody (mAb) candidates. By using a Golden Gate cloning (GGC)-dependent workflow, the bulk transfer of genetic information can be performed from antibody fragments displayed on yeast cells to a bidirectional mammalian expression vector. Herein, we describe in-depth protocols for the reformatting of mAbs, starting from the generation of Fab fragment libraries in YSD vectors and ending up with IgG molecules in bidirectional mammalian vectors in a consolidated two-pot, two-step procedure.

Keywords: Antibody hit discovery; Antibody reformatting; Golden Gate Cloning; Yeast surface display.

PubMed Disclaimer

Similar articles

See all similar articles

References

    1. Kaplon H, Chenoweth A, Crescioli S, Reichert JM (2022) Antibodies to watch in 2022. MAbs 14. https://doi.org/10.1080/19420862.2021.2014296
    1. Mullard A (2021) FDA approves 100th monoclonal antibody product. Nat Rev Drug Discov 20:491–495. https://doi.org/10.1038/d41573-021-00079-7 - DOI - PubMed
    1. Schaffitzel C, Hanes J, Jermutus L, Plückthun A (1999) Ribosome display: an in vitro method for selection and evolution of antibodies from libraries. J Immunol Methods 231:119–135. https://doi.org/10.1016/S0022-1759(99)00149-0 - DOI - PubMed
    1. McCafferty J, Griffiths AD, Winter G, Chiswell DJ (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554. https://doi.org/10.1038/348552a0 - DOI - PubMed
    1. Josephson K, Ricardo A, Szostak JW (2014) mRNA display: from basic principles to macrocycle drug discovery. Drug Discov Today 19:388–399. https://doi.org/10.1016/j.drudis.2013.10.011 - DOI - PubMed

LinkOut - more resources