Deep mutational engineering of broadly-neutralizing nanobodies accommodating SARS-CoV-1 and 2 antigenic drift

mAbs

卷:14

Laurent Bellanger¹  Fabrice Gallais¹  Stéphanie Debroas¹  Steven Dubois²  Raphaël Sierocki²  Adrien Laroche²  Hervé Nozach²  Bernard Maillère²  Benjamin Chalopin²  Stéphanie Simon³  Maria Lucia Orsini Delgado³  Philippe Cuniasse⁴ 

[1] CEA, INRAE, Département Médicaments et Technologies pour la Santé (DMTS), SPI, Université Paris Saclay, Bagnols-sur-Cèze, France;

[2] CEA, INRAE, Medicines and Healthcare Technologies Department, SIMoS, Université Paris-Saclay, Gif-sur-Yvette, France;

[3] CEA, INRAE, Medicines and Healthcare Technologies Department, SPI, Université Paris-Saclay, Gif-sur-Yvette, France;

[4] CNRS, CEA, Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, Gif-sur-Yvette, France;

关键词: Antibody engineering;    nanobodies;    deep mutational scanning;    yeast surface display;    SARS-CoV-2;   

DOI  :  10.1080/19420862.2022.2076775

来源: DOAJ

【 摘 要 】

Here, we report the molecular engineering of nanobodies that bind with picomolar affinity to both SARS-CoV-1 and SARS-CoV-2 receptor-binding domains (RBD) and are highly neutralizing. We applied deep mutational engineering to VHH72, a nanobody initially specific for SARS-CoV-1 RBD with little cross-reactivity to SARS-CoV-2 antigen. We first identified all the individual VHH substitutions that increase binding to SARS-CoV-2 RBD and then screened highly focused combinatorial libraries to isolate engineered nanobodies with improved properties. The corresponding VHH-Fc molecules show high affinities for SARS-CoV-2 antigens from various emerging variants and SARS-CoV-1, block the interaction between ACE2 and RBD, and neutralize the virus with high efficiency. Its rare specificity across sarbecovirus relies on its peculiar epitope outside the immunodominant regions. The engineered nanobodies share a common motif of three amino acids, which contribute to the broad specificity of recognition. Our results show that deep mutational engineering is a very powerful method, especially to rapidly adapt existing antibodies to new variants of pathogens.

【 授权许可】

Unknown