CDR1 composition can affect nanobody recombinant expression yields

Orlando, Marco (avtor), Fortuna, Sara (avtor), Oloketuyi, Sandra (avtor), Bajc, Gregor (avtor), Goldenzweig, Adi (avtor), De Marco, Ario (avtor)

The isolation of nanobodies from pre-immune libraries by means of biopanning is a straightforward process. Nevertheless, the recovered candidates often require optimization to improve some of their biophysical characteristics. In principle, CDRs are not mutated because they are likely to be part of the antibody paratope, but in this work, we describe a mutagenesis strategy that specifically addresses CDR1. Its sequence was identified as an instability hot spot by the PROSS program, and the available structural information indicated that four CDR1 residues bound directly to the antigen. We therefore modified the loop flexibility with the addition of an extra glycine rather than by mutating single amino acids. This approach significantly increased the nanobody yields but traded-off with moderate affinity loss. Accurate modeling coupled with atomistic molecular dynamics simulations enabled the modifications induced by the glycine insertion and the rationale behind the engineering design to be described in detail.

• Jezik: Angleški jezik
• Ključne besede: nanobodies, in silico engineering, mutagenesis, nanobody CDRs, in silico modeling, rational mutagenesis, nanobody engineering
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2021
• Št. strani: 13 str.
• Številčenje: Vol. 11, iss. 9, art. 1362
• PID: 20.500.12556/RUL-136353
• UDK: 577
• ISSN pri članku: 2218-273X
• DOI: 10.3390/biom11091362
• COBISS.SI-ID: 76397059
• Datum objave v RUL: 25.04.2022

Gradivo je del revije

• Naslov: Biomolecules
• Skrajšan naslov: Biomolecules
• Založnik: MDPI
• ISSN: 2218-273X
• COBISS.SI-ID: 519952921

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.
• Začetek licenciranja: 14.09.2021