Characterization of Fc?RIa (CD64) as a ligand molecule for site-specific IgG1 capture: A side-by-side comparison with protein a
| dc.authorid | 0000-0002-1677-644X | |
| dc.contributor.author | Çapkın, Eda | |
| dc.contributor.author | Kurt, Hasan | |
| dc.contributor.author | Gürel, Büşra | |
| dc.contributor.author | Bıçak, Dilan | |
| dc.contributor.author | Akgün Baş, Sibel | |
| dc.contributor.author | Dağlıkoca, Duygu Emine | |
| dc.contributor.author | Yüce, Meral | |
| dc.date.accessioned | 2023-02-22T06:19:36Z | |
| dc.date.available | 2023-02-22T06:19:36Z | |
| dc.date.issued | 2022 | |
| dc.department | İstanbul Medipol Üniversitesi, Mühendislik ve Doğa Bilimleri Fakültesi, Biyomedikal Mühendisliği Bölümü | |
| dc.department | İstanbul Medipol Üniversitesi, Rektörlük, Sağlık Bilim ve Teknolojileri Araştırma Enstitüsü | |
| dc.description.abstract | Fc ?receptors (Fc?Rs) are one of the structures that can initiate effector function for monoclonal antibodies. Fc?RIa has the highest affinity toward IgG1-type monoclonal antibodies among all Fc?Rs. In this study, a comprehensive characterization was performed for Fc?RIa as a potential affinity ligand for IgG1-type monoclonal antibody binding. The binding interactions were assessed with the SPR technique using different immobilization techniques such as EDC-NHS coupling, streptavidin-biotin interaction, and His-tagged Fc?RIa capture. The His-tagged Fc?RIa capture was the most convenient method based on assay repeatability. Next, a crude IgG1 sample and its fractions with different monomer contents obtained from protein A affinity chromatography were used to evaluate Fc?RIa protein in terms of monoclonal antibody binding capacity. The samples were also compared with a protein A-immobilized chip (a frequently used affinity ligand) for IgG1 binding responses. The antibody binding capacity of the protein A-immobilized chip surface was significantly better than that of the Fc?RIa-immobilized chip surface due to its 5 Ig binding domains. The antibody binding responses changed similarly with protein A depending on the monomer content of the sample. Finally, a different configuration was used to assess the binding affinity of free Fc?Rs (Fc?RIa, Fc?RIIa, and Fc?RIIIa) to three different immobilized IgGs by immobilizing protein L to the chip surface. Unlike previous immobilization techniques tested where the Fc?RIa was utilized as a ligand, nonimmobilized or free Fc?RIa resulted in a significantly higher antibody binding response than free protein A. In this configuration, kinetics data of Fc?RI revealed that the association rate (ka 50-80 × 105 M-1 s-1) increased in comparison to His capture method (1.9-2.4 × 105 M-1 s-1). In addition, the dissociation rate (kd 10-5 s-1) seemed slower over the His capture method (10-4 s-1) and provided stability on the chip surface during the dissociation phase. The KD values for Fc?RIa were found in the picomolar range (2.1-10.33 pM from steady-state affinity analysis and 37.5-46.2 pM from kinetic analysis) for IgG1-type antibodies. Fc?RIa possesses comparable ligand potential as well as protein A. Even though the protein A-immobilized surface bound more antibodies than the Fc?RIa-captured surface, Fc?RIa presented a significant antibody binding capacity in protein L configuration. The results suggest Fc?RIa protein as a potential ligand for site-oriented immobilization of IgG1-type monoclonal antibodies, and it needs further performance investigation on different surfaces and interfaces for applications such as sensing and antibody purification. | |
| dc.identifier.citation | Çapkın, E., Kurt, H., Gürel, B., Bıçak, D., Akgün Baş, S., Dağlıkoca, D. E. ... Yüce, M. (2022). Characterization of Fc?RIa (CD64) as a ligand molecule for site-specific IgG1 capture: A side-by-side comparison with protein a. Langmuir, 38(48), 14623-14634. https://doi.org/10.1021/acs.langmuir.2c02022 | |
| dc.identifier.doi | 10.1021/acs.langmuir.2c02022 | |
| dc.identifier.endpage | 14634 | |
| dc.identifier.issn | 0743-7463 | |
| dc.identifier.issn | 1520-5827 | |
| dc.identifier.issue | 48 | |
| dc.identifier.pmid | 36416530 | |
| dc.identifier.scopus | 2-s2.0-85143056054 | |
| dc.identifier.scopusquality | Q1 | |
| dc.identifier.startpage | 14623 | |
| dc.identifier.uri | https://doi.org/10.1021/acs.langmuir.2c02022 | |
| dc.identifier.uri | https://hdl.handle.net/20.500.12511/10497 | |
| dc.identifier.volume | 38 | |
| dc.identifier.wos | 000890684500001 | en_US |
| dc.identifier.wosquality | Q2 | |
| dc.indekslendigikaynak | Web of Science | |
| dc.indekslendigikaynak | Scopus | |
| dc.indekslendigikaynak | PubMed | |
| dc.institutionauthor | Kurt, Hasan | |
| dc.language.iso | en | |
| dc.publisher | American Chemical Society | |
| dc.relation.ispartof | Langmuir | en_US |
| dc.relation.publicationcategory | Makale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı | |
| dc.rights | Attribution 4.0 International | * |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | * |
| dc.subject | Fc?RIa (CD64) | |
| dc.subject | Specific IgG1 | |
| dc.subject | Protein A | |
| dc.title | Characterization of Fc?RIa (CD64) as a ligand molecule for site-specific IgG1 capture: A side-by-side comparison with protein a | |
| dc.type | Article |
