Ultrahigh-potent and broadly neutralizing anti-CD4 trimeric nanobodies inhibit HIV-1 infection by inducing CD4 conformational alteration
Xilin WuLinjing ZhuXiangyao WangFengfeng NiMingjun AoRuoke WangBin ZhengChen ChenLianfeng ShiShengya GengJia‐Qian HuMengshi YangDoudou ZhangPing YangMiaomiao LiYun‐Cheng LiQinxue HuSheng YePeng ZhengHongxia WeiLinqi ZhangYalan LiuZhiwei Wu
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Drug Development
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The neutralization of a constant dose of Semliki Forest virus by various doses of different antisera was studied. The presence of complement (1/200, v/v) in the neutralization mixture inhibited the neutralization by high concentration of antiserum and somewhat potentiated neutralization by low serum concentration. Because the experiments were biased towards a measure of irreversible neutralization, the inhibition observed in the presence of complement appeared to be an inhibition of irreversible neutralization. This inhibition was interpreted as a dissociation of a complement binding virus–antibody complex. The antibodies involved appeared to be virus-specific.
Semliki Forest virus
Dissociation constant
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The controlling characteristics of neutralization system in HPO process hexanolactam installation are presented. The reforming status of neutralization system and the methods for increasing the efficiency of neutralization and separation are described. [
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Objective:Assess cytotoxicity of medical device with silver nanoparticles and assay the reason of the cytotoxicity.Method:The nine kinds of silver nanoparticle samples in three types were tested by MTT cytotoxicity method.Result:The test showed that the cytotoxic ity in 9 final samples all were grade 3 to 4;the gels and dressings which are semi-finished products without silver nanoparticles,were grade 1 of cytotoxicity;patches were grade 4 of cytotoxicity;patches without adhesive were grade 1 of cytotoxicity.Conclusion:Silver nanoparticle medical devices in the present test showed moderate or severe cytotoxicity.Gels or dressings only which without silver nanoparticle showed slightly cytotoxicity.Patches only which without silver nanoparticle showed severe cytotoxicity,and adhesive is the main reason of the cytotoxicity.Whether the severe cytotoxicity will induce persist cell damage,and the detail mechanisms of silver nanoparticle or adhesive-induced cytotoxicity need further investigation.
Silver nanoparticle
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Broadly neutralizing antibodies (bNAbs) against HIV can reduce viral transmission in humans, but an effective therapeutic will require unusually high breadth and potency of neutralization. We employ the OSPREY computational protein design software to engineer variants of two apex-directed bNAbs, PGT145 and PG9RSH, resulting in increases in potency of over 100-fold against some viruses. The top designed variants improve neutralization breadth from 39% to 54% at clinically relevant concentrations (IC
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Examination was made of changes in the properties of N-Lauroyl-L-glutamate (LGA) with neutralization. Foaming power was maximum at neutralization of 1.6, and emulsification capacity decreased with neutralization. 13C-NMR indicated α-carboxylic acid of LGA to be neutralized predominantly, and γ-carboxylic acid to be so in proportion to neutralization at 1.6. Fluorescence probe application showed micropolarity and aggregation number of micelles to increase rapidly at neutralization of 1.6. α-carboxylic acid of LGA would thus appear to contribute to interactions between LGA molecules.
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Eight aza-chalcones L1-8 and eight new platinum(IV) complexes C1-8 with aza-chalcones were synthesized. The complexes have been characterized by elemental anaylsis, IR spectra, electronic spectrum and 1H NMR. The cytotoxicity was tested byMTT assays and compared with the cytotoxicity of cisplatinum. The results indicate that the complexes C1-3 and C5-8 have cytotoxicity against tested A-549 cell line; moreover, the cytotoxicity of complexes C6 approached cisplatinum. The complexes C3, C4 and C6 have cytotoxicity against tested Hela cell line, but the cytotoxicity of these complexes is lower than cisplatinum.Most of the coorespounding ligands have no cytotoxicity against tested two cell lines, and the cytotoxicity of all ligands is lower than complexes.The results suggest that the cytotoxicity of all complexes againstA-549 cell line is better than against Hela cell line.
HeLa
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The ELISA we developed was able to determine the antigen content and was suitable for a potency test, and we described a relative potency assay method which determines the potency of test vaccines by comparing the ELISA value of a test vaccine to that of a reference vaccine. In the present study, we standardized the reference vaccine used for determining the potencies of test vaccines, and established a potency test by ELISA. We evaluated the proposed reference vaccine by the neutralizing antibody responses in dogs after vaccination, by the challenge protection test in guinea pigs (GP potency test), which is the earlier official potency test used in Japan, and by the NIH potency test, which is widely used throughout the world. The results showed that a 4-fold dilution of the proposed reference vaccine induced sufficient immunity in dogs. A 3-fold dilution of the proposed reference vaccine passed the GP potency test. The international units (IU) calibrated by the NIH potency test were 3.7 IU/dose. From the results and the WHO recommendation that veterinary rabies vaccines should have a potency of at least 1.0 IU/dose, we determined to dilute the proposed reference vaccine by 3 fold and regarded it as the reference vaccine. Finally, we confirmed that there is a good agreement between the results of the potency test by ELISA and the results of the GP potency test. The establishment of the potency test by ELISA has made it possible to monitor the potency in the production process and has contributed to the stable production of the vaccine.
Rabies vaccine
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ABSTRACT Data are presented on the reliability of the previously described bioassay of gonadotrophin neutralizing potency of antisera raised against HCG preparations. The sensitivity of the method may be defined as the smallest detectable neutralizing activity expressed in anti-units (AU). This depends partly on the bioassay system used, and partly on the intended percentage of neutralization. In the HCG bioassay used in the present study, and around 50 % neutralization, the smallest detectable neutralizing activity is about 1.0 AU. The sensitivity decreases with increasing levels of neutralization: it is about 9 AU around 90% neutralization. The precision as given by the mean index of precision (λ) was 0.12 in 52 assays. The 95 % fiducial limits expressed as a percentage of the mean potency estimate ranged from 65.8 % to 125.7% in 52 assays conducted at 50 % and from 89.1% to 105.3% in 25 assays conducted at 90% intended neutralization. Reproducibility: eighteen antisera raised in 18 rabbits against 9 HCG preparations were assayed at 50% intended neutralization repeatedly (at least twice each antiserum) in 52 assays. Only one antiserum showed significant heterogeneity among repeated estimates by a χ2 test. The achieved neutralization ranged from 36 % to 79 % with a mean of 50 %. The neutralizing potency estimates of 19 antisera raised in 19 rabbits against 10 HCG preparations in 45 assays at 50% intended neutralization and in 25 assays at 90 % intended neutralization were also compared. These potency estimates were in close agreement with each other at both levels of neutralization (except for one antiserum) as evidenced by the results of χ 2 test and the value of the correlation coefficient obtained (r = 0.99). It is concluded that "the principle of simple additivity" is a satisfactory approximation to represent the antigen - antibody reaction involved in the neutralization of the biological activity of HCG preparations.
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Legal Cannabis products in the United States are required to report THC potency (total THC % by dry weight) on packaging, however concerns have been raised that reported THC potency values are inaccurate. Multiple studies have demonstrated that THC potency is a primary factor in determining pricing for Cannabis flower, so it has an outsized role in the marketplace. Reports of inflated THC potency and “lab shopping” to obtain higher THC potency results have been circulating for some time, but a side-by-side investigation of the reported potency and flower in the package has not previously been conducted. Using HPLC, we analyzed THC potency in 23 samples from 10 dispensaries throughout the Colorado Front Range and compared the results to the THC potency reported on the packaging. Average observed THC potency was 14.98 +/- 2.23%, which is substantially lower than recent reports summarizing dispensary reported THC potency. The average observed THC potency was 23.1% lower than the lowest label reported values and 35.6% lower than the highest label reported values. Overall, ~70% of the samples were more than 15% lower than the THC potency numbers reported on the label, with three samples having only one half of the reported maximum THC potency. Although the exact source of the discrepancies is difficult to determine, a lack of standardized testing protocols, limited regulatory oversight, and financial incentives to market high THC potency likely play a significant role. Given our results it is urgent that steps are taken to increase label accuracy of Cannabis being sold to the public. The lack of accurate reporting of THC potency can have impacts on medical patients controlling dosage, recreational consumers expecting an effect aligned with price, and trust in the industry as a whole. As the legal cannabis market continues to grow, it is essential that the industry moves toward selling products with more accurate labeling.
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