title: Thermal stability and refolding capability of shark derived single domain antibodies.
abstract: Single-domain antibodies (sdAb) from camelids and sharks represent the smallest immunoglobulin-based functional binding domains, and are known for their thermal stability and ability to refold after denaturation. Whereas target-binding sdAb have been derived from both immunized and naïve sharks and camelids, the stability of camelid-derived sdAb have been evaluated much more extensively. To address this disparity we characterized 20 sdAb derived from spiny dogfish shark and smooth dogfish shark in terms of their protein production, melting temperature and ability to refold after heat denaturation. Using the same expression system and protocol as we follow to produce camelid sdAb, production of the shark sdAb was quite poor, often resulting in less than a tenth of the typical yield for camelid sdAb. We measured the melting temperature of each of the sdAb. Similar to camelid sdAb, the shark-derived sdAb, showed a range of melting temperature values from 42°C to 77°C. Also similar to what has been observed in camelids, the sdAb from both shark species showed a range of ability to refold after heat denaturation. This work demonstrated that although shark sdAb can possess high melting temperatures and refolding ability, no clear advantage over sdAb derived from camelids in terms of thermostability and renaturation was obtained.
authors.affiliation: Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, 4555 Overlook Avenue, Washington, DC 20375, United States.
authors.forename: Jinny L
authors.initials: JL
authors.lastname: Liu
authors.affiliation: Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, 4555 Overlook Avenue, Washington, DC 20375, United States.
authors.forename: Dan
authors.initials: D
authors.lastname: Zabetakis
authors.affiliation: Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, 4555 Overlook Avenue, Washington, DC 20375, United States.
authors.forename: Jazmine C
authors.initials: JC
authors.lastname: Brown
authors.affiliation: Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, 4555 Overlook Avenue, Washington, DC 20375, United States.
authors.forename: George P
authors.initials: GP
authors.lastname: Anderson
authors.affiliation: Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, 4555 Overlook Avenue, Washington, DC 20375, United States. Electronic address: ellen.goldman@nrl.navy.mil.
authors.forename: Ellen R
authors.initials: ER
authors.lastname: Goldman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002161:Camelids, New World / Q000276:immunology; D006358:Hot Temperature; D008969:Molecular Sequence Data; D019151:Peptide Library; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D061905:Single-Domain Antibodies / Q000737:chemistry*; D048251:Squalus acanthias / Q000276:immunology*; D044366:Transition Temperature
chemical_list: D019151:Peptide Library; D061905:Single-Domain Antibodies
keywords: Antibody; Melting temperature; Smooth dogfish shark; Spiny dogfish shark; Stability