Rapid Antigen and Antibody-Like Molecule Discovery by Staphylococcal Surface Display

  • Marco CavallariEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2070)


Ever since the discovery of antibodies, they have been generated by complicated multi-step procedures. Typically, these involve sequencing, cloning, and screening after expression of the antibodies in a suitable organism and format. Here, a staphylococcal nanobody display is described that omits many the abovementioned intermediate steps and allows for simultaneous screening of multiple targets without prior knowledge nor expression of the binders. This paper reports a detailed, general step-by-step protocol to achieve nanobodies of high affinity. Apart from its focus on radioactive and fluorescent targets, it gives options for various other target formats and additional applications for the staphylococcal library; including flow cytometry and immunoprecipitation. This provides a system for antibody engineers that can be easily adopted to their specific needs.

Key words

Nanobody (VHH) Bacterial surface display Target identification Staphylococcal sortase A (SrtA) Immunoprecipitation 



Bovine serum albumin






Ethylenediaminetetraacetic acid


Green fluorescent protein


2-(N-morpholino)ethanesulfonic acid


Monosodium glutamate


Optical density


Phosphate-buffered saline


Room temperature


Sodium dodecyl sulfate




Tryptic soy agar


Tryptic soy broth


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.BIOSS Centre for Biological Signalling StudiesUniversity of FreiburgFreiburgGermany

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