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- 저자 : Hinterdorfer, P. (검색결과 3 건)
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Jin Caiquan,이채은,황현주,김예린,Hinterdorfer Peter,Myung Soon Chul,박성수,김미경,Hong Mineui,Ko Kisung 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.6
Multiple myeloma (MM) is an incurable disease characterized by malignant plasma cells within the bone marrow, and its increasing occurrence has highlighted the need for innovative strategies to address relapse and treatment resistance. Given the substantial expression of programmed death ligand 1 (PD-L1) in the human multiple myeloma cell line RPMI8226, we propose PD-L1 as a promising target for multiple myeloma therapy. Here, we successfully engineered an anti-PD-L1 monoclonal antibody (mAb) within a plant-based system. Building upon our previous findings, we germinated seeds derived from transgenic plants under in vitro conditions. Afterward, we screened the resulting seedlings for expression of the anti-PD-L1 mAb using polymerase chain reaction (PCR) and western blot analyses. Anti-PD-L1 mAbs were successfully purified from plant leaves and characterized through SDS-PAGE analysis. Our findings, which were confirmed via indirect enzyme-linked immunosorbent assay (ELISA), validate the binding affinity of the anti-PD-L1 mAb to recombinant PD-L1 protein. Furthermore, we investigated the interaction between the plant-derived anti-PD-L1 mAb and Fc gamma receptor I (FcγRI) as well as Fc gamma receptor IIIa (FcγRIIIa) molecules, confirming robust affinity. Additionally, the antibody’s binding affinity to the human multiple myeloma cancer cell line RPMI8226 was confirmed via cell ELISA. Our findings demonstrated that, unlike existing therapeutics, the plant-derived anti-PD-L1 antibody not only effectively binds to human recombinant PD-L1 protein but also to FcγRI and FcγRIIIa. These findings suggest the potential of plant-derived anti-PD-L1 mAb for the development of innovative therapies against multiple myeloma, emphasizing the need for further research and preclinical evaluation.
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Single-Molecule AFM Studies of Substrate Transport by Using the Sodium-Glucose Cotransporter SGLT1
Theeraporn Puntheeranurak,Rolf K. H. Kinne,Hermann J. Gruber,Peter Hinterdorfer 한국물리학회 2008 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.52 No.5
In an apical membrane of epithelial cells from the small intestine and the kidney, the high-affinity Na+/D-glucose cotransporter type 1 (SGLT1) plays a crucial role in intestinal glucose absorp- tion and in renal glucose reabsorption. Here, the over-expression of rabbit SGLT1 in rbSGLT1- transfected Chinese hamster ovary (CHO) cells was first characterized using the immuno-staining method on non-permeabilized cells. The cells were then imaged with atomic force microscopy (AFM), revealing live and fixed cells strongly attached to the glass surfaces. A bioconjugate chem- istry approach was employed to functionalize the surfaces of the AFM tips with D-glucose molecules via three dierent heterobifunctional crosslinkers. The D-glucose binding site and the translocation pathway of SGLT1 were investigated by studying the interaction forces between tip-bound D-glucose and SGLT1 in live cells on the single-molecule level. Analysis of these forces suggested that a long crosslinker with a small end group might be suitable for probing the D-glucose transport pathway of SGLT1. We show that single-molecule AFM technology is a powerful method for investigating transmembrane proteins and transporter functions in live cells. In an apical membrane of epithelial cells from the small intestine and the kidney, the high-affinity Na+/D-glucose cotransporter type 1 (SGLT1) plays a crucial role in intestinal glucose absorp- tion and in renal glucose reabsorption. Here, the over-expression of rabbit SGLT1 in rbSGLT1- transfected Chinese hamster ovary (CHO) cells was first characterized using the immuno-staining method on non-permeabilized cells. The cells were then imaged with atomic force microscopy (AFM), revealing live and fixed cells strongly attached to the glass surfaces. A bioconjugate chem- istry approach was employed to functionalize the surfaces of the AFM tips with D-glucose molecules via three dierent heterobifunctional crosslinkers. The D-glucose binding site and the translocation pathway of SGLT1 were investigated by studying the interaction forces between tip-bound D-glucose and SGLT1 in live cells on the single-molecule level. Analysis of these forces suggested that a long crosslinker with a small end group might be suitable for probing the D-glucose transport pathway of SGLT1. We show that single-molecule AFM technology is a powerful method for investigating transmembrane proteins and transporter functions in live cells.
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Characterization of Curli A Production on Living Bacterial Surfaces by Scanning Probe Microscopy
Oh, Y.,Cui, Y.,Kim, H.,Li, Y.,Hinterdorfer, P.,Park, S. Biophysical Society ; Published for the Biophysica 2012 Biophysical journal Vol.103 No.8
Curli are adhesive surface fibers produced by many Enterobacteriaceae, such as Escherichia coli and Salmonella enterica. They are implicated in bacterial attachment and invasion to epithelial cells. In this study, atomic force microscopy was used to determine the effects of curli on topology and mechanical properties of live E. coli cells. Young's moduli of both curli-deficient and curli-overproducing mutants were significantly lower than that of their wild-type (WT) strain, while decay lengths of the former strains were higher than that of the latter strain. Surprisingly, topological images showed that, unlike the WT and curli-overproducing mutant, the curli-deficient mutant produced a large number of flagella-like fibers, which may explain why the strain had a lower Young's modulus than the WT. These results suggest that the mechanical properties of bacterial surfaces are greatly affected by the presence of filamentous structures such as curli and flagella.
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