United Kingdom (UK)
Recombinant monoclonal antibody to RNA hairpin. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the phage display antibody BL3-6S97N.
Immunogen: A library of Fabs was generated by rationally varying the composition of the residues in complementarity determining regions (CDRs) of Fab BL3–6. Subsequently, the Fab library was challenged in a phage display selection to recognize and bind the GAAACAC motif.
Specificity: The antibody binds to the RNA hairpins GAAACAC hairpin. An RNA hairpin is an essential secondary structure of RNA. It can guide RNA folding, determine interactions in a ribozyme, protect messenger RNA (mRNA) from degradation, serve as a recognition motif for RNA binding proteins or act as a substrate for enzymatic reactions.
Application Notes: The Fab was generated by directed mutagenesis from the parental Fab BL3–6. The S97N mutation in complimentarity determining region L3 of the Fab improved the binding affinity compared to the parent Fab BL3–6 (Kd= 5 nM). The Fab fragment also bound to the affinity matured GAGACCC hairpin. The crystal structure of the Spinach RNA grafted with GAAACAC loop in complex with Fab BL3–6S97N was determined (Koirala et al., 2018; PMID: 29309709).
Antibody first published in:
Koirala et al. Affinity maturation of a portable Fab–RNA module for chaperone-assisted RNA crystallography Nucleic Acids Res. 2018 Mar 16;46(5):2624-2635. PMID:29309709
Note on publication:
The original paper describes the use of Fab fragment as chaperone for RNA crystallography.