PMID- 36416530
OWN - NLM
STAT- MEDLINE
DCOM- 20221207
LR  - 20230106
IS  - 1520-5827 (Electronic)
IS  - 0743-7463 (Print)
IS  - 0743-7463 (Linking)
VI  - 38
IP  - 48
DP  - 2022 Dec 6
TI  - Characterization of FcgammaRIa (CD64) as a Ligand Molecule for Site-Specific IgG1 
      Capture: A Side-By-Side Comparison with Protein A.
PG  - 14623-14634
LID - 10.1021/acs.langmuir.2c02022 [doi]
AB  - Fc gamma receptors (FcgammaRs) are one of the structures that can initiate effector 
      function for monoclonal antibodies. FcgammaRIa has the highest affinity toward 
      IgG1-type monoclonal antibodies among all FcgammaRs. In this study, a comprehensive 
      characterization was performed for FcgammaRIa 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 FcgammaRIa capture. The 
      His-tagged FcgammaRIa 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 
      FcgammaRIa 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 
      FcgammaRIa-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 FcgammaRs (FcgammaRIa, FcgammaRIIa, and FcgammaRIIIa) to three different 
      immobilized IgGs by immobilizing protein L to the chip surface. Unlike previous 
      immobilization techniques tested where the FcgammaRIa was utilized as a ligand, 
      nonimmobilized or free FcgammaRIa resulted in a significantly higher antibody binding 
      response than free protein A. In this configuration, kinetics data of FcgammaRI 
      revealed that the association rate (k(a) 50-80 x 10(5) M(-1) s(-1)) increased in 
      comparison to His capture method (1.9-2.4 x 10(5) M(-1) s(-1)). In addition, the 
      dissociation rate (k(d) 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 K(D) values for FcgammaRIa 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. FcgammaRIa possesses comparable ligand potential as well as 
      protein A. Even though the protein A-immobilized surface bound more antibodies 
      than the FcgammaRIa-captured surface, FcgammaRIa presented a significant antibody binding 
      capacity in protein L configuration. The results suggest FcgammaRIa 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.
FAU - Capkin, Eda
AU  - Capkin E
AD  - Faculty of Engineering and Natural Sciences, Sabanci University, Tuzla 34956, 
      Istanbul, Turkey.
FAU - Kurt, Hasan
AU  - Kurt H
AD  - School of Engineering and Natural Sciences, Istanbul Medipol University, Beykoz 
      34810, Istanbul, Turkey.
AD  - SABITA Research Institute for Health Sciences and Technologies, Istanbul Medipol 
      University, Beykoz 34810, Istanbul, Turkey.
AD  - Nanosolar Plasmonics Ltd., Gebze 41400, Kocaeli, Turkey.
FAU - Gurel, Busra
AU  - Gurel B
AD  - SUNUM Nanotechnology Research and Application Center, Sabanci University, Tuzla 
      34956, Istanbul, Turkey.
FAU - Bicak, Dilan
AU  - Bicak D
AD  - ILKO ARGEM Biotechnology R&D Center, Pendik 34906, Istanbul, Turkey.
FAU - Akgun Bas, Sibel
AU  - Akgun Bas S
AD  - ILKO ARGEM Biotechnology R&D Center, Pendik 34906, Istanbul, Turkey.
FAU - Daglikoca, Duygu Emine
AU  - Daglikoca DE
AD  - ILKO ARGEM Biotechnology R&D Center, Pendik 34906, Istanbul, Turkey.
FAU - Yuce, Meral
AU  - Yuce M
AUID- ORCID: 0000-0003-0393-1225
AD  - SUNUM Nanotechnology Research and Application Center, Sabanci University, Tuzla 
      34956, Istanbul, Turkey.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221123
PL  - United States
TA  - Langmuir
JT  - Langmuir : the ACS journal of surfaces and colloids
JID - 9882736
RN  - 0 (Receptors, IgG)
RN  - 0 (Staphylococcal Protein A)
RN  - 0 (Immunoglobulin G)
RN  - 0 (Ligands)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antibodies, Immobilized)
SB  - IM
MH  - *Receptors, IgG/chemistry/metabolism
MH  - *Staphylococcal Protein A/chemistry/metabolism
MH  - Immunoglobulin G/chemistry
MH  - Ligands
MH  - Kinetics
MH  - Antibodies, Monoclonal
MH  - Antibodies, Immobilized
MH  - Protein Binding
PMC - PMC9730901
COIS- The authors declare no competing financial interest.
EDAT- 2022/11/24 06:00
MHDA- 2022/12/10 06:00
PMCR- 2022/12/08
CRDT- 2022/11/23 08:53
PHST- 2022/11/24 06:00 [pubmed]
PHST- 2022/12/10 06:00 [medline]
PHST- 2022/11/23 08:53 [entrez]
PHST- 2022/12/08 00:00 [pmc-release]
AID - 10.1021/acs.langmuir.2c02022 [doi]
PST - ppublish
SO  - Langmuir. 2022 Dec 6;38(48):14623-14634. doi: 10.1021/acs.langmuir.2c02022. Epub 
      2022 Nov 23.