Easily Established and Multifunctional Synthetic Nanobody Libraries as Research Tools

Liu, Bing-Ying; Yang, Daiwen

doi:10.3390/ijms23031482

Cited by 24 publications

(29 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immune libraries are the most effective approach for obtaining functional nanobodies. As the Figure 1 shows, to construct immune libraries, Camillidae animals, like dromedary, alpaca, llama and Heavy Chain Only Antibodies (HCAbs) transgenic mice are actively immunized with a special immunogen (Janssens et al, 2006;Liu and Yang, 2022). These immunogens stimulate the immune system to raise HCAbs with reasonable specificity and high affinity (De Genst et al, 2004).…”

Section: Construction Of Nanobody Librarymentioning

confidence: 99%

“…Liu and Yang summarized a systematic approach to construct synthetic libraries, including the selection of suitable framework sequences, the design of CDRs' randomization, and the synthesis of a sufficient number of DNAs with different sequences. The undesirable frameshift mutation can be controlled through optimizing the primer quality, the DNA polymerase fidelity, and experimental techniques, in order to improve the quality of synthetic libraries (Liu and Yang, 2022).…”

Section: Construction Of Nanobody Librarymentioning

confidence: 99%

“…Phage display technique based on E. coli expression systems is the most common screening method. Fv genes are constructed into the phage genome and displayed on the phage surface together with phage coat protein (Liu and Yang, 2022). Phages are secreted by E.…”

Section: Expression System Of Nanobodymentioning

confidence: 99%

See 2 more Smart Citations

Research progress and applications of nanobody in human infectious diseases

et al. 2022

View full text Add to dashboard Cite

Infectious diseases, caused by pathogenic microorganisms, are capable of affecting crises. In addition to persistent infectious diseases such as malaria and dengue fever, the vicious outbreaks of infectious diseases such as Neocon, Ebola and SARS-CoV-2 in recent years have prompted the search for more efficient and convenient means for better diagnosis and treatment. Antibodies have attracted a lot of attention due to their good structural characteristics and applications. Nanobodies are the smallest functional single-domain antibodies known to be able to bind stably to antigens, with the advantages of high stability, high hydrophilicity, and easy expression and modification. They can directly target antigen epitopes or be constructed as multivalent nanobodies or nanobody fusion proteins to exert therapeutic effects. This paper focuses on the construction methods and potential functions of nanobodies, outlines the progress of their research, and highlights their various applications in human infectious diseases.

show abstract

Section: Construction Of Nanobody Librarymentioning

confidence: 99%

Section: Construction Of Nanobody Librarymentioning

confidence: 99%

See 1 more Smart Citation

Research progress and applications of nanobody in human infectious diseases

et al. 2022

View full text Add to dashboard Cite

show abstract

“…Considering the restricted application of antibodies in some fields, NBs could be regarded as a promising alternative for them 50 . The small size, high stability, and remarkable specificity of NBs make them promising candidates for biological applications 51 . One of the most attractive uses of NBs is cancer therapy 52 , which could be applied to various cancer cells.…”

Section: Discussionmentioning

confidence: 99%

Modeling and affinity maturation of an anti-CD20 nanobody: a comprehensive in-silico investigation

Poustforoosh

Faramarz

Negahdaripour

et al. 2023

Sci Rep

View full text Add to dashboard Cite

B-cell Non-Hodgkin lymphomas are the malignancies of lymphocytes. CD20 is a membrane protein, which is highly expressed on the cell surface of the B-cells in NHL. Treatments using monoclonal antibodies (mAbs) have resulted in failure in some cases. Nanobodies (NBs), single-domain antibodies with low molecular weights and a high specificity in antigen recognition, could be practical alternatives for traditional mAbs with superior characteristics. To design an optimized NB as a candidate CD20 inhibitor with raised binding affinity to CD20, the structure of anti-CD20 NB was optimized to selectively target CD20. The 3D structure of the NB was constructed based on the optimal templates (6C5W and 5JQH), and the key residues were determined by applying a molecular docking study. After identifying the key residues, some mutations were introduced using a rational protocol to improve the binding affinity of the NB to CD20. The rational mutations were conducted using the experimental design (Taguchi method). Six residues (Ser27, Thr28, Phe29, Ile31, Asp99, and Asn100) were selected as the key residues, and five residues were targeted for rational mutation (Trp, Phe, His, Asp, and Tyr). Based on the mutations suggested by the experimental design, two optimized NB structures were constructed. NB2 showed a remarkable binding affinity to CD20 in docking studies with a binding energy of − 853 kcal/mol. The optimized NB was further evaluated using molecular dynamics simulation. The results revealed that CDR1 (complementarity determining regions1) and CDR3 are essential loops for recognizing the antigen. NB2 could be considered as a potential inhibitor of CD20, though experimental evaluations are needed to confirm it.

show abstract

“…More recently, synthetically generated nanobody libraries (which utilise a nanobody-based scaffold with synthetically generated diversity within the binding regions) have been developed ( Cabanillas-Bernal et al, 2019 ; Moutel et al, 2016 ; Zimmermann et al, 2018 ; reviewed in Liu and Yang, 2022 ). Engineering of the nanobody scaffold in synthetic libraries can be exploited to improve biochemical stability and ease of protein folding (e.g.…”

Section: Affimer and Nanobody Screening And Applicationsmentioning

confidence: 99%

Affimers and nanobodies as molecular probes and their applications in imaging

Cordell

Carrington

Curd

et al. 2022

Journal of Cell Science

View full text Add to dashboard Cite

Antibodies are the most widely used, traditional tool for labelling molecules in cells. In the past five to ten years, many new labelling tools have been developed with significant advantages over the traditional antibody. Here, we focus on nanobodies and the non-antibody binding scaffold proteins called Affimers. We explain how they are generated, selected and produced, and we describe how their small size, high binding affinity and specificity provides them with many advantages compared to antibodies. Of particular importance, their small size enables them to better penetrate dense cytoskeletal regions within cells, as well as tissues, providing them with specific advantage for super-resolution imaging, as they place the fluorophore with a few nanometres of the target protein being imaged. We expect these novel tools to be of broad interest to many cell biologists and anticipate them becoming the tools of choice for super-resolution imaging.

show abstract

Easily Established and Multifunctional Synthetic Nanobody Libraries as Research Tools

Cited by 24 publications

References 62 publications

Research progress and applications of nanobody in human infectious diseases

Research progress and applications of nanobody in human infectious diseases

Modeling and affinity maturation of an anti-CD20 nanobody: a comprehensive in-silico investigation

Affimers and nanobodies as molecular probes and their applications in imaging

Contact Info

Product

Resources

About