Tunable pharmacokinetics: modifying the in vivo half-life of antibodies by directed mutagenesis of the Fc fragment

Immunoglobulins (Igs) are large proteins of 150 kDa with prolonged residence time in blood. Their half-life is controlled by their ability to interact with the protective neonatal Fc receptor (FcRn, Brambell receptor) present on endothelial cells. Here, we describe a protocol using site-specific mutagenesis of individual residues responsible for this interaction, resulting in engineered antibodies with distinct half-lives. The method is a powerful tool that enables manipulation of half-lives and is applicable to all antibodies and Fc fusion proteins for the development of agents with controlled pharmacokinetic properties. Moreover, the protocol is applicable to any situation where the structure and/or function of engineered proteins are to be studied. The protocol begins with the mutagenesis reaction at the DNA level and proceeds to describe mammalian expression and purification of recombinant proteins, radiolabeling and evaluation in vivo. The time frame for completing the procedure is about 6 months, provided that no complications are encountered.     

Tunable promoters in systems biology

The construction of synthetic promoter libraries has represented a major breakthrough in systems biology, enabling the subtle tuning of enzyme activities. A number of tools are now available that allow the modulation of gene expression and the detection of changes in expression patterns. But, how does one choose the correct promoter and what are the appropriate methods for reading promoter strength? Furthermore, how fine should the tuning of gene expression be for some specific applications and how can the simultaneous and individual tuning of multiple genes be achieved? Some recent studies have helped us to find answers to many of these questions.     

Polarization-Controlled Tunable Multi-focal Plasmonic Lens

A polarization-controlled tunable plasmonic lens which can generate different multi-focal combinations with exciting sources of left and right circular polarizations is proposed in this paper. Both position and intensity of each focal point can be adjusted by modulating the structure of the plasmonic lens. It is believed that the polarization-controlled tunable plasmonic multi-focal lens can be potentially used for optical switches and multi-channel couplers in future logic photonic and plasmonic systems.     

Tunable plasma relativistic microwave amplifier

A stable regime of the amplification of a slow plasma wave in a plasma waveguide during the injection of a high-current relativistic electron beam is obtained. For an input-signal frequency of 9.1 GHz, there exists a range of plasma densities in which the spectrum of the output microwave radiation lies in a 0.5-GHz-wide band. For a 40-kW input power at a frequency of 9.1 GHz, the maximum output power is 8 MW. It is shown experimentally for the first time that the beam-plasma amplifier can operate at frequencies of 9.1 GHz and 12.9 GHz. The range of plasma densities in which the regime of amplification is observed agrees with the results of calculations based on linear theory.     

Tunable Spoof Surface Plasmons Bulleye Antenna

A tunable spoof surface plasmons antenna using sinusoidally modulated corrugated reactance surface based on a bulleye structure is proposed in this paper. The designed antenna is made of concentric metallic grooves etched on a metal plate, the depth of which is of sinusoidal periodic variation in the radial direction. This makes it possible that highly confined spoof surface plasmons along corrugated surface can be converted to radiation modes. The proposed bulleye antenna can work from 25.8 to 33 GHz and a bandwidth of 7.2 GHz and its main lobe can be directed at 30^∘ from the vertical direction at 30 GHz. This antenna has a maximum gain of 15 dB and its main lobe can scan from 14^∘ to 58^∘ by tuning the frequency from 28 to 32 GHz.

     

Flexible and Tunable Dielectric Color Meta-hologram

Plasmonics - Metasurface holograms consisting of nanostructures have shown great promise for various applications due to their unique capability of shaping light. However, most of previous work...     

Tunable Plasmonic Nanolaser Based on Graphene

Plasmonics - Surface plasmon polariton nanolaser, which can achieve all-optical circuits and optoelectronic integration, is a major research area in nano-optics. We propose a novel tunable...     

Tunable Assembly of Peptide-coated Gold Nanoparticles

The interaction between peptides and gold surfaces has increasingly been of interest for bionanotechnology applications. To more fully understand how to control such interactions, we have studied the optical properties of peptide-modified gold nanoparticles as a function of peptide composition, pH of the surrounding medium, and peptide concentration. We show using localized surface plasmon resonance, transmission electron microscopy, and surface-enhanced Raman scattering (SERS) that selected “gold-binding peptides” (GBPs), similar to those isolated for binding to gold films using yeast display, can bind to gold nanoparticles at a variety of pHs. Peptide modifications of nanoparticles can lead to irreversible particle aggregation when the pH of the solution is kept below the isoelectric point (pI) of the peptide. However, at pHs above the peptide’s pI, particles remain stable in solution, and peptides remain bound to the particles possibly through amine coordination of gold. Additionally, we demonstrate the potential in using SERS for the direct detection of GBPs on gold-silica nanoshells, eliminating the need for indirect labeling methods.     

Tunable cell-surface mimetics as engineered cell substrates

Most recent breakthroughs in understanding cell adhesion, cell migration, and cellular mechanosensitivity have been made possible by the development of engineered cell substrates of well-defined surface properties. Traditionally, these substrates mimic the extracellular matrix (ECM) environment by the use of ligand-functionalized polymeric gels of adjustable stiffness. However, such ECM mimetics are limited in their ability to replicate the rich dynamics found at cell-cell contacts. This review focuses on the application of cell surface mimetics, which are better suited for the analysis of cell adhesion, cell migration, and cellular mechanosensitivity across cell-cell interfaces. Functionalized supported lipid bilayer systems were first introduced as biomembrane-mimicking substrates to study processes of adhesion maturation during adhesion of functionalized vesicles (cell-free assay) and plated cells. However, while able to capture adhesion processes, the fluid lipid bilayer of such a relatively simple planar model membrane prevents adhering cells from transducing contractile forces to the underlying solid, making studies of cell migration and cellular mechanosensitivity largely impractical. Therefore, the main focus of this review is on polymer-tethered lipid bilayer architectures as biomembrane-mimicking cell substrate. Unlike supported lipid bilayers, these polymer-lipid composite materials enable the free assembly of linkers into linker clusters at cellular contacts without hindering cell spreading and migration and allow the controlled regulation of mechanical properties, enabling studies of cellular mechanosensitivity. The various polymer-tethered lipid bilayer architectures and their complementary properties as cell substrates are discussed.     

Tunable Dielectric Properties of Ferrite-Dielectric Based Metamaterial

A ferrite-dielectric metamaterial composed of dielectric and ferrite cuboids has been investigated by experiments and simulations. By interacting with the electromagnetic wave, the Mie resonance can take place in the dielectric cuboids and the ferromagnetic precession will appear in the ferrite cuboids. The magnetic field distributions show the electric Mie resonance of the dielectric cuboids can be influenced by the ferromagnetic precession of ferrite cuboids when a certain magnetic field is applied. The effective permittivity of the metamaterial can be tuned by modifying the applied magnetic field. A good agreement between experimental and simulated results is demonstrated, which confirms that these metamaterials can be used for tunable microwave devices.     

A Tunable Coarse-Grained Model for Ligand-Receptor Interaction

Cell-surface receptors are the most common target for therapeutic drugs. The design and optimization of next generation synthetic drugs require a detailed understanding of the interaction with their corresponding receptors. Mathematical approximations to study ligand-receptor systems based on reaction kinetics strongly simplify the spatial constraints of the interaction, while full atomistic ligand-receptor models do not allow for a statistical many-particle analysis, due to their high computational requirements. Here we present a generic coarse-grained model for ligand-receptor systems that accounts for the essential spatial characteristics of the interaction, while allowing statistical analysis. The model captures the main features of ligand-receptor kinetics, such as diffusion dependence of affinity and dissociation rates. Our model is used to characterize chimeric compounds, designed to take advantage of the receptor over-expression phenotype of certain diseases to selectively target unhealthy cells. Molecular dynamics simulations of chimeric ligands are used to study how selectivity can be optimized based on receptor abundance, ligand-receptor affinity and length of the linker between both ligand subunits. Overall, this coarse-grained model is a useful approximation in the study of systems with complex ligand-receptor interactions or spatial constraints.     

Surface-Enhanced Raman Scattering in Tunable Bimetallic Core-Shell

In this paper, optical properties of multilayer spherical core-shell nanoparticles based on quasi-static approach and plasmon hybridization theory are investigated. Calculations show that light absorption spectrum of bimetallic multilayer core-shell has three intense plasmon resonance peaks, which are more suitable for multiplex biosensing based on surface-enhanced Raman scattering (SERS) and localized surface plasmon resonance (LSPR). The plasmon resonance peaks in bimetal nanshells are optimized by tuning the geometrical parameters. In addition, the optimal geometry is discussed to obtain the Raman enhancement factor in bimetallic multilayer nanoshell. SERS enhancement factor is calculated with consideration of dampings due to both the electron scattering and the radiation at the boundary and modified Drude model in dielectric function of bimetallic nanoshell. It is shown that bimetallic nanoshell with the small size exhibits strong SERS enhancement factor (~6.63 × 10⁵) with additional collision dampings and ~2.9 × 10⁹ with modified Drude model which are suitable for biosensing applications. In addition, any variation in blood concentration and oxygen level can be detected by this bimetallic core-shell nanoparticle with sensitivity of Δλ/Δn = 264.91 nm/RIU.     

Tunable self-assembly of genetically engineered silk--elastin-like protein polymers

Silk--elastin-like protein polymers (SELPs), consisting of the repeating units of silk and elastin blocks, combine a set of outstanding physical and biological properties of silk and elastin. Because of the unique properties, SELPs have been widely fabricated into various materials for the applications in drug delivery and tissue engineering. However, little is known about the fundamental self-assembly characteristics of these remarkable polymers. Here we propose a two-step self-assembly process of SELPs in aqueous solution for the first time and report the importance of the ratio of silk-to-elastin blocks in a SELP's repeating unit on the assembly of the SELP. Through precise tuning of the ratio of silk to elastin, various structures including nanoparticles, hydrogels, and nanofibers could be generated either reversibly or irreversibly. This assembly process might provide opportunities to generate innovative smart materials for biosensors, tissue engineering, and drug delivery. Furthermore, the newly developed SELPs in this study may be potentially useful as biomaterials for controlled drug delivery and biomedical engineering.     

Tunable Extraordinary THz Transmission Using Liquid Metal-Based Devices

In this paper, we report novel designs of tunable THz plasmonic devices based on liquid metals. The designed devices will be able to dynamically control and change the spectrum responses of extraordinary THz wave transmissions. Different THz device configurations are investigated, and numerical simulations have been conducted to theoretically verify the performance of the proposed structures. Moreover, an equivalent circuit model has been developed to describe the operating principle of the proposed THz devices. Good agreement has been achieved between the theoretical models and the numerical results. These new THz devices are expected to be applied in various areas of sensing, communication, and imaging.     

Tunable degradation of maleimide-thiol adducts in reducing environments

Addition chemistries are widely used in preparing biological conjugates, and in particular, maleimide-thiol adducts have been widely employed. Here, we show that the resulting succinimide thioether formed by the Michael-type addition of thiols to N-ethylmaleimide (NEM), generally accepted as stable, undergoes retro and exchange reactions in the presence of other thiol compounds at physiological pH and temperature, offering a novel strategy for controlled release. Model studies ((1)H NMR, HPLC) of NEM conjugated to 4-mercaptophenylacetic acid (MPA), N-acetylcysteine, or 3-mercaptopropionic acid (MP) incubated with glutathione showed half-lives of conversion from 20 to 80 h, with extents of conversion from 20% to 90% for MPA and N-acetylcysteine conjugates. After ring-opening, the resultant succinimide thioether did not show retro and exchange reactions. The kinetics of the retro reactions and extent of exchange can be modulated by the Michael donor's reactivity; therefore, the degradation of maleimide-thiol adducts could be tuned for controlled release of drugs or degradation of materials at time scales different than those currently possible via disulfide-mediated release. Such approaches may find a new niche for controlled release in reducing environments relevant in chemotherapy and subcellular trafficking.     

Tunable signal processing in synthetic MAP kinase cascades

The flexibility of MAPK cascade responses enables regulation of a vast array of cell fate decisions, but elucidating the mechanisms underlying this plasticity is difficult in endogenous signaling networks. We constructed insulated mammalian MAPK cascades in yeast to explore how intrinsic and extrinsic perturbations affect the flexibility of these synthetic signaling modules. Contrary to biphasic dependence on scaffold concentration, we observe monotonic decreases in signal strength as scaffold concentration increases. We find that augmenting the concentration of sequential kinases can enhance ultrasensitivity and lower the activation threshold. Further, integrating negative regulation and concentration variation can decouple ultrasensitivity and threshold from the strength of the response. Computational analyses show that cascading can generate ultrasensitivity and that natural cascades with different kinase concentrations are innately biased toward their distinct activation profiles. This work demonstrates that tunable signal processing is inherent to minimal MAPK modules and elucidates principles for rational design of synthetic signaling systems.     

Tunable calcium current through TRPV1 receptor channels

TRPV1 receptors are polymodal cation channels that open in response to diverse stimuli including noxious heat, capsaicin, and protons. Because Ca2+ is vital for TRPV1 signaling, we sought to precisely measure its contribution to TRPV1 responses and discovered that the Ca2+ current was tuned by the mode of activation. Using patch clamp photometry, we found that the fraction of the total current carried by Ca2+ (called the Pf%) was significantly smaller for TRPV1 currents evoked by protons than for those evoked by capsaicin. Using site-directed mutagenesis, we discovered that the smaller Pf% was due to protonation of three acidic amino acids (Asp646, Glu648, and Glu651) that are located in the mouth of the pore. Thus, in keeping with recent reports of time-dependent changes in the ionic permeability of some ligand-gated ion channels, we now show for the first time that the physiologically important Ca2+ current of the TRPV1 receptor is also dynamic and depends on the mode of activation. This current is significantly smaller when the receptor is activated by a change in pH, owing to atomic scale interactions of H+ and Ca2+ with the fixed negative charge of side chains in the pore.     

Tunable Multi-Port Surface Plasmon Polariton Excitation with Nanostructures

Surface plasmon polaritons (SPPs) have appealing features such as tighter spatial confinement and higher local field intensity. Manipulation of surface plasmon polaritons on metal/dielectric interface is an important aspect in the achievement of integrated plasmonic circuit beyond the diffraction limit. Here, we introduce a design of pin cushion structure and a holographic groove pattern structure for tunable multi-port SPPs excitation and focusing. Free space light is coupled into SPPs through momentum matching conditions. Both nanostructures are capable of tunably controlling of SPPs depending on the incident polarizations, while the holographic method provides more flexibility of wavelength-dependent excitations. Furthermore, a quantitative method is applied to calculate the efficiencies of excitation for both nanostructures under different conditions, including radially polarized incident beams. These results can work as a guidance and be helpful to further choice of the suitable design strategies for variable plasmonic applications such as beam splitter, on-chip spectroscopy, and plasmonic detectors.     

A Tunable Ultra-Broadband Metamaterial Absorber with Multilayered Structure

In this article, we demonstrate a tunable ultra-broadband metamaterial absorber (TUMA) in terahertz (THz) band which is based on the multilayered structure composed of an Au reflective layer, polyimide dielectric layers, and vanadium dioxide (VO₂) periodic structures, respectively. We gain the tunable absorption spectra because of the room temperature phased-changed character of VO2. The relative bandwidth reaches to 81.2% and the absorption rate is over 90% at the frequency range of 1.63–3.86 THz when the temperature (t₁) is 350 K, but when t₁ = 300 K, the presented absorber is acted as a reflector whose absorption is small besides the frequency points of 9.75 THz and 9.81 THz. For the sake of comprehending the physical mechanism in-depth, the electric field (E-field) diagrams, the surface current distributions and the power loss density (PLD) of the TUMA are investigated. The influences of structural arguments and incident angle (θ) on the absorption are also analyzed. The emulated consequences show that the absorption spectrum can be regulated by changing structural parameters and incident angle and the tunable absorption regions can be obtained by altering the external temperature.