333. Selection of Neutralizing Antibody-Resistant AAV8 Variants with Structure-Guided Site-Specific Saturated Mutagenesis

Nipah virus (NiV) represents a significant pandemic threat with zoonotic transmission from bats-to-humans with almost annual regional outbreaks characterized by documented human-to-human transmission and high fatality rates. Currently, no vaccine against NiV has been approved. Structure-based design and protein engineering principles were applied to stabilize the fusion (F) protein in its prefusion trimeric conformation (pre-F) to improve expression and increase immunogenicity. We covalently linked the stabilized pre-F through trimerization domains at the C-terminus to three attachment protein (G) monomers, forming a chimeric design. These studies detailed here focus on mRNA delivery of NiV immunogens in mice, assessment of mRNA immunogen-specific design elements and their effects on humoral and cellular immunogenicity. The pre-F/G chimera elicited a strong neutralizing antibody response and a superior NiV-specific Tfh and other effector T cell response compared to G alone across both the mRNA and protein platforms. These findings enabled final candidate selection of pre-F/G Fd for clinical development.

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Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs

Scientific Reports ◽

10.1038/srep13348 ◽

2015 ◽

Vol 5 (1) ◽

Cited By ~ 44

Author(s):

Xianlong Wang ◽

Jinwei Zhou ◽

Chunwei Cao ◽

Jiaojiao Huang ◽

Tang Hai ◽

...

Keyword(s):

Gene Disruption ◽

Site Specific ◽

Highly Active ◽

Selection Of

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Selection of site-specific regression model for characterization of uniaxial compressive strength of rock

International Journal of Rock Mechanics and Mining Sciences ◽

10.1016/j.ijrmms.2015.01.008 ◽

2015 ◽

Vol 75 ◽

pp. 73-81 ◽

Cited By ~ 57

Author(s):

Yu Wang ◽

Adeyemi Emman Aladejare

Keyword(s):

Compressive Strength ◽

Regression Model ◽

Uniaxial Compressive Strength ◽

Site Specific ◽

Selection Of

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Strategies for Monitoring Multiuse Trail Networks

Transportation Research Record Journal of the Transportation Research Board ◽

10.3141/2644-12 ◽

2017 ◽

Vol 2644 (1) ◽

pp. 100-110 ◽

Cited By ~ 2

Author(s):

Greg Lindsey ◽

Jeffrey S. Wilson ◽

Jueyu Wang ◽

Tracy Hadden-Loh

Keyword(s):

Nonprofit Organizations ◽

Large Scale ◽

Low Cost ◽

Facility Planning ◽

Monitoring Networks ◽

Site Specific ◽

New Challenges ◽

Large Scale Networks ◽

Selection Of

Many municipalities, park districts, and nonprofit organizations have begun monitoring nonmotorized traffic on multiuse trails as the need for information about the use of facilities has grown and relatively low-cost sensors for automated monitoring have become available. As they have gained experience, they have begun to move from site-specific monitoring on individual trails to a more comprehensive monitoring of trail networks. This case study review compares strategies developed by 10 organizations for monitoring traffic on multiuse trails, including local, multicounty, statewide, and multistate trail networks. The focus is on approaches to the design of monitoring networks, particularly the rationales or objectives for monitoring and the selection of monitoring sites. It is shown that jurisdictions are following principles of monitoring established by FHWA and that the design of monitoring networks is evolving to meet new challenges, including monitoring large-scale networks. Relevant outcomes and implications for practice are summarized. The researchers concluded that FHWA guidelines can be adapted to many circumstances and can increase information for decision making. Trail monitoring is informing decisions related to facility planning, investment, and safety.

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Economic Modeling of Site-Specific Optimum Wind Turbine for Electrification Studies

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.347-353.1973 ◽

2011 ◽

Vol 347-353 ◽

pp. 1973-1986 ◽

Cited By ~ 1

Author(s):

Umar Bawah ◽

Khaled E Addoweesh ◽

Ali M. Eltamaly

Keyword(s):

Wind Turbine ◽

Initial Study ◽

Capital Cost ◽

Economic Modeling ◽

Decision Makers ◽

Site Specific ◽

Initial Capital ◽

Cost Of Energy ◽

Economic Selection ◽

Selection Of

A generalized approach for the economic selection of wind turbine for a given wind regime is proposed in this paper. It draws from the literature and standards being used in the field to arrive at an economic site specific wind turbine based on minimizing the annual cost of energy produced (AEP) while tracking the initial capital cost (ICC) of investment required. It is meant to provide an initial study to guide decision makers who are contemplating using wind energy as a power source to generate electricity in commercial quantity for community usage. It is a general estimation approach which does not require surfing for manufacture prices and wind turbine parameters. The input data consists of site specific wind data, hub height, rotor diameter and turbine power rating. The output gives a range of plots of feasible wind turbine ratings, rotor diameters, rated speed against initial capital cost (ICC) and also cost of energy produced (COE).

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Chemical Cross-Linking Stabilizes Native-Like HIV-1 Envelope Glycoprotein Trimer Antigens

Journal of Virology ◽

10.1128/jvi.01942-15 ◽

2015 ◽

Vol 90 (2) ◽

pp. 813-828 ◽

Cited By ~ 21

Author(s):

Torben Schiffner ◽

Natalia de Val ◽

Rebecca A. Russell ◽

Steven W. de Taeye ◽

Alba Torrents de la Peña ◽

...

Keyword(s):

Envelope Glycoprotein ◽

Morphological Characteristics ◽

Neutralizing Antibody ◽

Structural Instability ◽

Cross Linking ◽

Public Health Importance ◽

Protective Antibodies ◽

Hiv 1 ◽

Chemical Cross Linking ◽

Selection Of

ABSTRACTMajor neutralizing antibody immune evasion strategies of the HIV-1 envelope glycoprotein (Env) trimer include conformational and structural instability. Stabilized soluble trimers such as BG505 SOSIP.664 mimic the structure of virion-associated Env but nevertheless sample different conformational states. Here we demonstrate that treating BG505 SOSIP.664 trimers with glutaraldehyde or a heterobifunctional cross-linker introduces additional stability with relatively modest effects on antigenicity. Thus, most broadly neutralizing antibody (bNAb) epitopes were preserved after cross-linking, whereas the binding of most weakly or nonneutralizing antibodies (non-NAb) was reduced. Cross-linking stabilized all Env conformers present within a mixed population, and individual conformers could be isolated by bNAb affinity chromatography. Both positive selection of cross-linked conformers using the quaternary epitope-specific bNAbs PGT145, PGT151, and 3BC315 and negative selection with non-NAbs against the V3 region enriched for trimer populations with improved antigenicity for bNAbs. Similar results were obtained using the clade B B41 SOSIP.664 trimer. The cross-linking method may, therefore, be useful for countering the natural conformational heterogeneity of some HIV-1 Env proteins and, by extrapolation, also vaccine immunogens from other pathogens.IMPORTANCEThe development of a vaccine to induce protective antibodies against HIV-1 is of primary public health importance. Recent advances in immunogen design have provided soluble recombinant envelope glycoprotein trimers with near-native morphology and antigenicity. However, these trimers are conformationally flexible, potentially reducing B-cell recognition of neutralizing antibody epitopes. Here we show that chemical cross-linking increases trimer stability, reducing binding of nonneutralizing antibodies while largely maintaining neutralizing antibody binding. Cross-linking followed by positive or negative antibody affinity selection of individual stable conformational variants further improved the antigenic and morphological characteristics of the trimers. This approach may be generally applicable to HIV-1 Env and also to other conformationally flexible pathogen antigens.

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Probing neutralizing-antibody responses against emerging measles viruses (MVs): immune selection of MV by H protein-specific antibodies?

Journal of General Virology ◽

10.1099/vir.0.80467-0 ◽

2005 ◽

Vol 86 (2) ◽

pp. 365-374 ◽

Cited By ~ 37

Author(s):

Sabine Santibanez ◽

Stefan Niewiesk ◽

Alla Heider ◽

Jürgen Schneider-Schaulies ◽

Guy A. M. Berbers ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Measles Virus ◽

Protein A ◽

Neutralizing Antibody ◽

Antibody Responses ◽

Neutralizing Capacity ◽

Human Sera ◽

H Protein ◽

Neutralizing Epitopes ◽

Selection Of

Measles virus (MV) infection and vaccination induce long-lasting immunity and neutralizing-antibody responses that are directed against the MV haemagglutinin (H) and the fusion (F) protein. A new MV genotype, D7, emerged recently in western Germany and rapidly replaced the long-term endemically circulating genotypes C2 and D6. Analysis of the H gene of C2, D6, D7 and vaccine viruses revealed uniform sequences for each genotype. Interestingly, a consistent exchange of seven distinct amino acids in the D7 H was observed when compared with residues shared between C2, D6 and vaccine viruses, and one exchange (D416→N) in the D7 H was associated with an additional N-linked glycosylation. In contrast, the F gene is highly conserved between MVs of these genotypes. To test whether the D7 H protein escapes from antibody responses that were raised against earlier circulating or vaccine viruses, the neutralizing capacity of mAbs recognizing seven distinct domains on the H of an Edmonston-related MV was compared. The mAbs revealed a selective and complete loss of two neutralizing epitopes on the D7 H when compared with C2, D6 and vaccine viruses. To assess whether these alterations of the D7 H affect the neutralizing capacity of polyclonal B-cell responses, genotype-specific antisera were produced in cotton rats. However, no significant genotype-dependent difference was found. Likewise, human sera obtained from vaccinees (n=7) and convalescents (n=6) did not distinguish between the MV genotypes. Although the hypothesis of selection of D7 viruses by pre-existing neutralizing antibodies is compatible with the differing pattern of neutralizing epitopes on the H protein, it was not confirmed by the results of MV neutralization with polyclonal sera.

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