Bionanocapsule-based enzyme–antibody conjugates for enzyme-linked immunosorbent assay

Iijima, Masumi; Matsuzaki, Takashi; Kadoya, Hiroyasu; Hatahira, Satoko; Hiramatsu, Shingo; Jung, Giman; Tanizawa, Katsuyuki; Kuroda, Shinya

doi:10.1016/j.ab.2009.10.010

Cited by 26 publications

(21 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this study, we could directly observe real-time movement of mouse IgG3s tethered onto ZZ-BNCs in solution by HS-AFM. It was therefore confirmed that ZZ-BNCs achieved oriented immobilization of IgGs, which has been hypothesized by functional analyses91011, by tethering Fc subunits in nearly close-packed arrangement and minimizing the steric hindrance of Fv regions for efficient antigen bindings. Since ZZ-BNCs are stable to heat, chemical, and mechanical stresses9, ZZ-BNCs would be an ideal and practical scaffold for oriented immobilization of IgGs.…”

Section: Discussionsupporting

confidence: 61%

“…ZZ-BNC contains about 120 molecules of ZZ-L protein (N-terminally ZZ-fused L protein) embedded in a liposome and has ability for capturing ~60 mouse IgG molecules, as well as displaying all the IgG Fv regions outwardly9. Furthermore, ZZ-BNCs can enhance the sensitivity of immunosensors9 and immunoassays1011 not only through the oriented immobilization of antibodies but also the clustering of antibodies and labelling molecules. Thus, ZZ-BNC is a promising scaffold for a variety of conventional immunosensors and immunoassays.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Nano-visualization of oriented-immobilized IgGs on immunosensors by high-speed atomic force microscopy

Iijima

Somiya

Yoshimoto

et al. 2012

Sci Rep

Self Cite

View full text Add to dashboard Cite

Oriented immobilization of sensing molecules on solid phases is an important issue in biosensing. In case of immunosensors, it is essential to scrutinize not only the direction and shape of immunoglobulin G (IgG) in solution but also the real-time movement of IgGs, which cannot be achieved by conventional techniques. Recently, we developed bio-nanocapsules (BNCs) displaying a tandem form of the IgG Fc-binding Z domain derived from Staphylococcus aureus protein A (ZZ-BNC) to enhance the sensitivity and antigen-binding capacity of IgG via oriented-immobilization. Here, we used high-speed atomic force microscopy (HS-AFM) to reveal the fine surface structure of ZZ-BNC and observe the movement of mouse IgG3 molecules tethered onto ZZ-BNC in solution. ZZ-BNC was shown to act as a scaffold for oriented immobilization of IgG, enabling its Fv regions to undergo rotational Brownian motion. Thus, HS-AFM could decipher real-time movement of sensing molecules on biosensors at the single molecule level.

show abstract

Section: Discussionsupporting

confidence: 61%

mentioning

confidence: 99%

Nano-visualization of oriented-immobilized IgGs on immunosensors by high-speed atomic force microscopy

Iijima

Somiya

Yoshimoto

et al. 2012

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…The silicon surface was cleaned with oxygen plasma in a plasma asher (200 W, 5 min; JPA300, J-science) and treated with 1 % HF for 1 min. After repeating the plasma (10 min) and 1 % HF treatment once again, the nanoneedle was modified by physical adsorption of 50 µg/ml of ZZ-BNC at room temperature for 1 h; ZZ-BNC is bio-nano capsule based anchor to which Fc domain of antibodies can bind [27–29]. Anti-vimentin antibody (V6630, Sigma-Aldrich) was bound to the ZZ-domain of the ZZ-BNC by incubating with 475 µg/ml antibody in PBS at room temperature for 1 h. The antibody-immobilized nanoneedle was rinsed in prior to the use in cell fishing experiments.…”

Section: Methodsmentioning

confidence: 99%

High efficiency penetration of antibody-immobilized nanoneedle thorough plasma membrane for in situ detection of cytoskeletal proteins in living cells

et al. 2016

View full text Add to dashboard Cite

BackgroundThe field of structural dynamics of cytoskeletons in living cells is gathering wide interest, since better understanding of cytoskeleton intracellular organization will provide us with not only insights into basic cell biology but may also enable development of new strategies in regenerative medicine and cancer therapy, fields in which cytoskeleton-dependent dynamics play a pivotal role. The nanoneedle technology is a powerful tool allowing for intracellular investigations, as it can be directly inserted into live cells by penetrating through the plasma membrane causing minimal damage to cells, under the precise manipulation using atomic force microscope. Modifications of the nanoneedles using antibodies have allowed for accurate mechanical detection of various cytoskeletal components, including actin, microtubules and intermediate filaments. However, successful penetration of the nanoneedle through the plasma membrane has been shown to vary greatly between different cell types and conditions. In an effort to overcome this problem and improve the success rate of nanoneedle insertion into the live cells, we have focused here on the fluidity of the membrane lipid bilayer, which may hinder nanoneedle penetration into the cytosolic environment.ResultsWe aimed to reduce apparent fluidity of the membrane by either increasing the approach velocity or reducing experimental temperatures. Although changes in approach velocity did not have much effect, lowering the temperature was found to greatly improve the detection of unbinding forces, suggesting that alteration in the plasma membrane fluidity led to increase in nanoneedle penetration.ConclusionsOperation at a lower temperature of 4 °C greatly improved the success rate of nanoneedle insertion to live cells at an optimized approach velocity, while it did not affect the binding of antibodies immobilized on the nanoneedle to vimentins for mechanical detection. As these experimental parameters can be applied to various cell types, these results may improve the versatility of the nanoneedle technology to other cell lines and platforms.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-016-0226-5) contains supplementary material, which is available to authorized users.

show abstract

“…BNCs and ZZ-BNCs were purified as described previously. 18,26 Protein concentrations were determined with a bicinchoninic acid (BCA) protein assay kit (Pierce, Rockford, IL) using bovine serum albumin (BSA; Wako Pure Chemical Industries, Osaka, Japan) as a control protein. BNC was labeled with a Fluorolink Cy5 monofunctional reactive dye (GE Healthcare, Milwaukee, WI) and CF750 NHS ester (Biotium, Heyward, CA) according to the manufacturer's protocol.…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

“…13 These results demonstrated that the α-CD11c-ZZ-BNC complexes could more promptly and efficiently accumulate in splenic DCs (in 62% of CD11c + splenocytes at 40 minutes after intravenous injection) than other DC-targeting nanocarriers. Because ZZBNCs could display antibodies in an oriented-immobilization manner and thereby significantly enhance the sensitivity, antigen-binding capacity, and affinity of the antibody itself, 24,26,27 complex formation by α-CD11c antibodies with ZZ-BNCs may improve the molecular recognition between CD11c molecules and DC-targeting nanocarriers.…”

Section: Next Cd11cmentioning

confidence: 99%

Engineered hepatitis B virus surface antigen L protein particles for in vivo active targeting of splenic dendritic cells

Matsuo

Yoshimoto²,

Iijima³

et al. 2012

IJN

Self Cite

View full text Add to dashboard Cite

Dendritic cells (DCs) are key regulators of adaptive T-cell responses. By capturing exogenous antigens and presenting antigen-derived peptides via major histocompatibility complex molecules to naïve T cells, DCs induce antigen-specific immune responses in vivo. In order to induce effective host immune responses, active delivery of exogenous antigens to DCs is considered important for future vaccine development. We recently generated bionanocapsules (BNCs) consisting of hepatitis B virus surface antigens that mediate stringent in vivo cell targeting and efficient endosomal escape, and after the fusion with liposomes (LP) containing therapeutic materials, the BNC-LP complexes deliver them to human liver-derived tissues in vivo. BNCs were further modified to present the immunoglobulin G (IgG) Fc-interacting domain (Z domain) derived from Staphylococcus aureus protein A in tandem. When mixed with IgGs, modified BNCs (ZZ-BNCs) displayed the IgG Fv regions outwardly for efficient binding to antigens in an oriented-immobilization manner. Due to the affinity of the displayed IgGs, the IgG-ZZ-BNC complexes accumulated in specific cells and tissues in vitro and in vivo. After mixing ZZ-BNCs with antibodies against DCs, we used immunocytochemistry to examine which antibodies delivered ZZ-BNCs to mouse splenic DCs following intravenous injection of the ZZ-BNCs. ZZ-BNCs displaying anti-CD11c monoclonal antibodies (α-CD11c-ZZ-BNCs) were found to accumulate with approximately 62% of splenic DCs, and reside within some of them. After the fusion with liposomes containing antigens, the α-CD11c-ZZ-BNCs could elicit the respective antibodies more efficiently than other nontargeting control vaccines, suggesting that this DC-specific nanocarrier is promising for future vaccines.

show abstract

Bionanocapsule-based enzyme–antibody conjugates for enzyme-linked immunosorbent assay

Cited by 26 publications

References 16 publications

Nano-visualization of oriented-immobilized IgGs on immunosensors by high-speed atomic force microscopy

Nano-visualization of oriented-immobilized IgGs on immunosensors by high-speed atomic force microscopy

High efficiency penetration of antibody-immobilized nanoneedle thorough plasma membrane for in situ detection of cytoskeletal proteins in living cells

Engineered hepatitis B virus surface antigen L protein particles for in vivo active targeting of splenic dendritic cells

Contact Info

Product

Resources

About