Our original paper on the development of first-generation nanoCLAMP scaffold: "Development of polyol-responsive antibody mimetics for single-step protein purification" in Protein Expr. Purif. 2017. Here we describe the development of the first antibody mimetics that can release bound protein upon exposure to propylene glycol or glycerol. nanoCLAMPs (nano CLostridial Antibody Mimetic Proteins) are based on a monomeric 16 kDa beta sandwich domain with loops analogous to the CDRs of the immunoglobulin variable domain.
Our recent paper on the engineering of nanoCLAMPs to resist heat, alkali, proteases and organic solvents: "Protein engineering of a nanoCLAMP antibody mimetic scaffold as a platform for producing bioprocess-compatible affinity capture ligands" in J. Biol. Chem. 2023. Here we describe the development of next-generation nanoCLAMPs that retain binding activity after exposure to heat (including autoclaving), 0.1 mg/ml trypsin, 0.1 M NaOH, and 100% DMF. The original scaffold was improved via 7 rounds of mutagenesis. The video below shows in green the 30 residues that have been mutated in the improved scaffold. The variable loops are shown in orange, and the remaining residues in gray.
Guo Z, Smutok O, Johnston WA, Walden P, Ungerer JPJ, Peat TS, Newman J, Parker J, Nebl T, Hepburn C, Melman A, Suderman RJ, Katz E, Alexandrov K. Design of a methotrexate-controlled chemical dimerization system and its use in bio-electronic devices. Nat Commun. 2021 Dec 8;12(1):7137. doi: 10.1038/s41467-021-27184-w. PMID: 34880210; PMCID: PMC8654847.
Quentin Pagneux, Nathalie Garnier, Manon Fabregue, Sarah Sharkaoui, Sophie Mazzoli, Ilka Engelmann, Rabah Boukherroub, Mary Strecker, Eric Cruz, Peter Ducos, Ana Zarubica, Richard Suderman, Sabine Szunerits. nanoCLAMP potently neutralizes SARS-CoV-2 and protects K18-hACE2 mice from infection. bioRxiv.2023.04.03. 535401;doi:https://doi.org/10.1101/2023.04.03.535401.