Title : Secretory production of a camelid single-domain antibody (VHH, nanobody) by the Serratia marcescens Lip system in Escherichia coli - Asada_2021_Biochem.Biophys.Res.Commun_549_105 |
Author(s) : Asada T , Takagi D , Nakai M , Abe S , Yuasa K |
Ref : Biochemical & Biophysical Research Communications , 549 :105 , 2021 |
Abstract :
Escherichia coli is one of the most popularly used hosts to produce recombinant proteins. Most recombinant proteins are produced in the cytoplasm and periplasm, requiring multiple steps to extract and purify recombinant proteins. The Serratia marcescens Lip system (LipB-LipC-LipD) is a type 1 secretion system that selectively secretes LipA from the intracellular to extracellular space in a single step. This study aimed to establish a secretory production system for nanobodies, camelid-derived small molecule antibody fragments, using the S. marcescens Lip system. Surprisingly, E. coli harboring only LipC, a membrane fusion protein of the Lip system, could secrete an anti-green fluorescent protein (GFP)-Nb, a nanobody against GFP, without the addition of a long amino acid sequence. The LipC-based secretion system recognized the Val-Thr-Val sequence at the C-terminus of the nanobody. Finally, Strep-tagged anti-GFP-Nb was purified from culture supernatants of E. coli harboring LipC by Strep-affinity chromatography at a final yield of >5 mg per liter of culture supernatant. These results potently supported that the S. marcescens LipC-based secretion system has the potential to establish an efficient secretory production system for nanobodies. |
PubMedSearch : Asada_2021_Biochem.Biophys.Res.Commun_549_105 |
PubMedID: 33667707 |
Asada T, Takagi D, Nakai M, Abe S, Yuasa K (2021)
Secretory production of a camelid single-domain antibody (VHH, nanobody) by the Serratia marcescens Lip system in Escherichia coli
Biochemical & Biophysical Research Communications
549 :105
Asada T, Takagi D, Nakai M, Abe S, Yuasa K (2021)
Biochemical & Biophysical Research Communications
549 :105