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CDR1 composition can affect nanobody recombinant expression yields
ID
Orlando, Marco
(
Author
),
ID
Fortuna, Sara
(
Author
),
ID
Oloketuyi, Sandra
(
Author
),
ID
Bajc, Gregor
(
Author
),
ID
Goldenzweig, Adi
(
Author
),
ID
De Marco, Ario
(
Author
)
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MD5: FE32011D99471EC24A7DDD657486D25D
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https://www.mdpi.com/2218-273X/11/9/1362
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Abstract
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.
Language:
English
Keywords:
nanobodies
,
in silico engineering
,
mutagenesis
,
nanobody CDRs
,
in silico modeling
,
rational mutagenesis
,
nanobody engineering
Work type:
Article
Typology:
1.01 - Original Scientific Article
Organization:
BF - Biotechnical Faculty
Publication status:
Published
Publication version:
Version of Record
Year:
2021
Number of pages:
13 str.
Numbering:
Vol. 11, iss. 9, art. 1362
PID:
20.500.12556/RUL-136353
UDC:
577
ISSN on article:
2218-273X
DOI:
10.3390/biom11091362
COBISS.SI-ID:
76397059
Publication date in RUL:
25.04.2022
Views:
950
Downloads:
94
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Record is a part of a journal
Title:
Biomolecules
Shortened title:
Biomolecules
Publisher:
MDPI
ISSN:
2218-273X
COBISS.SI-ID:
519952921
Licences
License:
CC BY 4.0, Creative Commons Attribution 4.0 International
Link:
http://creativecommons.org/licenses/by/4.0/
Description:
This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:
14.09.2021
Projects
Funder:
ARRS - Slovenian Research Agency
Project number:
N4-0046
Name:
Identifikacija rekombinantnih nanotelesc za imunsko detekcijo eksosomov za diagnozo raka na dojkah
Funder:
ARRS - Slovenian Research Agency
Project number:
J4-9322
Name:
Razvoj reagentov za diagnostično stratificiranje in tarčno zdravljenje raka na dojki na osnovi tekočinskih biopsij
Funder:
Other - Other funder or multiple funders
Funding programme:
Italian Association for Cancer Research (AIRC)
Project number:
IG 2020-ID.24589
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