Development of nanobodies specific to clumping factors A of Staphylococcus aureus by yeast surface display
Copyright © 2024 Elsevier B.V. All rights reserved..
The Staphylococcus aureus clumping factor A (ClfA) is a fibrinogen (Fg) binding protein that plays an important role in the clumping of S. aureus in blood plasma. The current anti-infective approaches targeting ClfA are mainly based on monoclonal antibodies but showed less impressive efficacy for clinical applications. Nanobodies offer advantages in enhanced tissue penetration and a propensity to bind small epitopes. However, there is no report on generating specific nanobodies for ClfA. Here, we constructed a synthetic nanobody library based on yeast surface display to isolate nanobodies against the Fg binding domain ClfA221-550. We firstly obtained a primary nanobody directed to ClfA221-550, and then employed error-prone mutagenesis to enhance its binding affinity. Finally, 18 variants were isolated with high affinities (EC50, 1.1 ± 0.1 nM to 4.8 ± 0.3 nM), in which CNb1 presented the highest inhibition efficiency in the adhesion of S. aureus to fibrinogen. Moreover, structural simulation analysis indicated that the epitope for CNb1 partially overlapped with the binding sites for fibrinogen, thus inhibiting ClfA binding to Fg. Overall, these results indicated that the specific nanobodies generated here could prevent the adhesion of S. aureus to fibrinogen, suggesting their potential capacities in the control of S. aureus infections.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:259 |
|---|---|
| Enthalten in: |
International journal of biological macromolecules - 259(2024), Pt 2 vom: 20. Feb., Seite 129208 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Mei, Meng [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
9001-32-5 |
|---|
| Anmerkungen: |
Date Completed 22.02.2024 Date Revised 22.02.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1016/j.ijbiomac.2024.129208 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM366759183 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM366759183 | ||
| 003 | DE-627 | ||
| 005 | 20240222232303.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 240114s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.ijbiomac.2024.129208 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1302.xml |
| 035 | |a (DE-627)NLM366759183 | ||
| 035 | |a (NLM)38185298 | ||
| 035 | |a (PII)S0141-8130(24)00011-4 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Mei, Meng |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Development of nanobodies specific to clumping factors A of Staphylococcus aureus by yeast surface display |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 22.02.2024 | ||
| 500 | |a Date Revised 22.02.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2024 Elsevier B.V. All rights reserved. | ||
| 520 | |a The Staphylococcus aureus clumping factor A (ClfA) is a fibrinogen (Fg) binding protein that plays an important role in the clumping of S. aureus in blood plasma. The current anti-infective approaches targeting ClfA are mainly based on monoclonal antibodies but showed less impressive efficacy for clinical applications. Nanobodies offer advantages in enhanced tissue penetration and a propensity to bind small epitopes. However, there is no report on generating specific nanobodies for ClfA. Here, we constructed a synthetic nanobody library based on yeast surface display to isolate nanobodies against the Fg binding domain ClfA221-550. We firstly obtained a primary nanobody directed to ClfA221-550, and then employed error-prone mutagenesis to enhance its binding affinity. Finally, 18 variants were isolated with high affinities (EC50, 1.1 ± 0.1 nM to 4.8 ± 0.3 nM), in which CNb1 presented the highest inhibition efficiency in the adhesion of S. aureus to fibrinogen. Moreover, structural simulation analysis indicated that the epitope for CNb1 partially overlapped with the binding sites for fibrinogen, thus inhibiting ClfA binding to Fg. Overall, these results indicated that the specific nanobodies generated here could prevent the adhesion of S. aureus to fibrinogen, suggesting their potential capacities in the control of S. aureus infections | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a ClfA | |
| 650 | 4 | |a Fibrinogen | |
| 650 | 4 | |a Nanobody | |
| 650 | 7 | |a Single-Domain Antibodies |2 NLM | |
| 650 | 7 | |a Fibrinogen |2 NLM | |
| 650 | 7 | |a 9001-32-5 |2 NLM | |
| 700 | 1 | |a Lu, Mengqing |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Shiqi |e verfasserin |4 aut | |
| 700 | 1 | |a Ren, Xinyi |e verfasserin |4 aut | |
| 700 | 1 | |a Xing, Banbin |e verfasserin |4 aut | |
| 700 | 1 | |a Hu, Yang |e verfasserin |4 aut | |
| 700 | 1 | |a Wu, Yuqi |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Huan |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Longhao |e verfasserin |4 aut | |
| 700 | 1 | |a Yi, Li |e verfasserin |4 aut | |
| 700 | 1 | |a Ming, Ke |e verfasserin |4 aut | |
| 700 | 1 | |a Wei, Zigong |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t International journal of biological macromolecules |d 1992 |g 259(2024), Pt 2 vom: 20. Feb., Seite 129208 |w (DE-627)NLM012627356 |x 1879-0003 |7 nnns |
| 773 | 1 | 8 | |g volume:259 |g year:2024 |g number:Pt 2 |g day:20 |g month:02 |g pages:129208 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.ijbiomac.2024.129208 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 259 |j 2024 |e Pt 2 |b 20 |c 02 |h 129208 | ||