全文获取类型
收费全文 | 427篇 |
免费 | 55篇 |
国内免费 | 11篇 |
出版年
2024年 | 2篇 |
2023年 | 5篇 |
2022年 | 6篇 |
2021年 | 16篇 |
2020年 | 18篇 |
2019年 | 23篇 |
2018年 | 23篇 |
2017年 | 28篇 |
2016年 | 17篇 |
2015年 | 24篇 |
2014年 | 20篇 |
2013年 | 39篇 |
2012年 | 23篇 |
2011年 | 19篇 |
2010年 | 17篇 |
2009年 | 15篇 |
2008年 | 10篇 |
2007年 | 24篇 |
2006年 | 12篇 |
2005年 | 6篇 |
2004年 | 15篇 |
2003年 | 15篇 |
2002年 | 18篇 |
2001年 | 5篇 |
2000年 | 3篇 |
1999年 | 6篇 |
1998年 | 4篇 |
1997年 | 8篇 |
1996年 | 4篇 |
1995年 | 6篇 |
1994年 | 3篇 |
1993年 | 8篇 |
1992年 | 5篇 |
1991年 | 3篇 |
1990年 | 3篇 |
1989年 | 4篇 |
1988年 | 4篇 |
1987年 | 3篇 |
1986年 | 1篇 |
1985年 | 7篇 |
1984年 | 2篇 |
1983年 | 2篇 |
1982年 | 6篇 |
1981年 | 1篇 |
1980年 | 3篇 |
1979年 | 4篇 |
1978年 | 1篇 |
1976年 | 1篇 |
1973年 | 1篇 |
排序方式: 共有493条查询结果,搜索用时 15 毫秒
191.
Although human IgG heavy chain genes encode a C‐terminal lysine, this residue is mostly absent from the endogenous antibodies isolated from serum. Some low but variable level of C‐terminal lysine is present on therapeutic antibodies expressed in mammalian cell culture systems. Here, we monitored the C‐terminal lysine processing of a recombinant human IgG2 antibody after intravenous injection into human subjects. Peptide mapping of the therapeutic antibody isolated from serum samples by affinity purification was used to quantify the C‐terminal lysine levels over time in vivo. The C‐terminal lysine residue was found to be rapidly lost in vivo with a half life of about an hour (62 min). In vivo C‐terminal lysine processing could be reproduced in vitro, but at a faster rate, by incubating in human serum. Pretreated serum, under conditions used to inactivate carboxypeptidase U, generated in vitro C‐terminal lysine processing rates that more closely matched those in vivo. Endogenous IgG, isolated from human blood, contained very low levels of C‐terminal lysine (~0.02%), consistent with the expected circulating half life of antibodies and the calculated C‐terminal lysine processing rate. Thus, the low residual IgG2 C‐terminal lysine is rapidly processed in vivo and such processing likely occurs on endogenous antibodies in circulation. Biotechnol. Bioeng. 2011;108: 404–412. © 2010 Wiley Periodicals, Inc. 相似文献
192.
Yuichi Takiue Makoto Hosoyamada Takuya Yokoo Masaki Kimura Manami Ochiai Kiyoko Kaneko 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):596-600
Familial juvenile hyperuricemic nephropathy is caused by mutations in the UMOD gene encoding uromodulin. A transgenic mouse model was developed by introducing a human mutant UMOD (C148W) cDNA under control of the mouse umod promoter. Uromodulin accumulation was observed in the thick ascending limb cells in the kidney of transgenic mice. However, the urinary excretion of uromodulin in transgenic mice did not decrease and LC-MS/MS analysis indicated it was of mouse origin. Moreover, the creatinine clearance was not different between wildtype and transgenic animals. Consequently, the onset of the disease was not observed in transgenic mice until 24 weeks of age. 相似文献
193.
The purpose of the present study was to evaluate the efficacies and mechanisms of the PAB (para-amino benzamidine) affinity
column chromatography, virus filtration, pasteurization (60°C heat treatment for 10 h), and lyophilization steps employed
in the manufacture of urokinase from human urine with regard to the removal and/or inactivation of human immunodeficiency
virus (HIV), bovine viral diarrhoea virus (BVDV), bovine herpes virus (BHV), and murine encephalomyocarditis virus (EMCV).
Samples from relevant stages of the production process were spiked with each virus and subjected to scale-down processes mimicking
the manufacture of urokinase. Samples were collected at each step, immediately titrated using a 50% tissue culture infectious
dose (TCID50), and the virus reduction factors evaluated. PAB chromatography was found to be an effective step for removing BVDV, BHV,
and EMCV with log reduction factors of 2.79, 6.50, and 5.96, respectively. HIV, BVDV, BHV, and EMCV were completely removed
during the Viresolve NFP filtration step with log reduction factors of ≥6.06, ≥4.60, ≥5.44, and ≥6.87, respectively. Pasteurization
was also found to be a robust and effective step in inactivating all the viruses tested, since there were no residual viruses
detected after the pasteurization process. The log reduction factors achieved by pasteurization were ≥5.73 for HIV, ≥3.86
for BVDV, ≥6.75 for BHV, and ≥5.92 for EMCV. Lyophilization showed significant efficacy for inactivating BVDV, BHV, and EMCV
with log reduction factors of 2.69, 1.37, and 4.70, respectively. These results indicate that the production process for urokinase
exhibited a sufficient viral reducing capacity to achieve a high margin of virus safety. 相似文献
194.
In the production of monoclonal antibodies (mAbs) intended for use in humans, it is a global regulatory requirement that the manufacturing process includes unit operations that are proven to inactivate or remove adventitious agents to ensure viral safety. Viral inactivation by low pH hold (LPH) is typically used to ensure this viral safety in the purification process of mAbs and other biotherapeutics derived from mammalian cell lines. To ascertain the effectiveness of the LPH step, viral clearance studies have evaluated LPH under worst-case conditions of pH above the manufacturing set point and hold duration at or below the manufacturing minimum. Highly acidic conditions (i.e., pH < 3.60) provide robust and effective enveloped virus inactivation but may lead to reduced product quality of the therapeutic protein. However, when viral inactivation is operated above pH 3.60 to ensure product stability, effective (>4 log10 reduction factor) viral inactivation may not be observed under these worst-case pH conditions in viral clearance studies. A multivariate design of experiments was conducted to further characterize the operating space for low pH viral inactivation of a model retrovirus, xenotropic murine leukemia virus (X-MuLV). The statistically designed experiment evaluated the effect of mAb isotype, pH, temperature, acid titrant, sodium chloride (NaCl) concentration, virus spike timing, and post-spike filtration on X-MuLV inactivation. Data from the characterization study were used to generate predictive models to identify conditions that reliably achieve effective viral inactivation at pH ≥ 3.60. Results of the study demonstrated that NaCl concentration has the greatest effect on virus inactivation in the range studied, and pH has a large effect when the load material has no additional NaCl. Overall, robust and effective inactivation of X-MuLV at pH 3.65–3.80 can be achieved by manipulating either the pH or the NaCl concentration of the load material. This study contributes to the understanding of ionic strength as an influential parameter in low pH viral inactivation studies. 相似文献
195.
The clearance rates (CRs) of bacteria by Filinia longiseta were measured at 19°C, both in situ in Lake Loosdrecht and in the laboratory during summer. The tracer particles used in the field were: (1) 0.51 µm fluorescent microspheres, and (2) fluorescently labelled bacteria (FLB). A third type of tracer particle, natural [methyl-3H]-thymidine-labelled bacteria (< 1.2 µm), were used as a radiotracer in a laboratory experiment. The uptake of the first two tracer-particle types was measured by microscopic examination of the rotifer guts. In the third case, the uptake of radioactivity was determined by liquid scintillation counting. The rate of uptake of the microspheres decreased 10 min after the start of the experiment, probably because the gut passage time was exceeded. Using a 5 min feeding time, the rate of uptake of microspheres was higher than that of the FLB, though the variation in the uptake in both cases was high. The ingestion rates and CRs of bacteria by F. longiseta based on the fluorescent tracers were: microspheres, 5115 bact.ind–1 h–1 and 0.368 µl ind–1 h–1; FLB, 2252 bact.ind–1 h–1 and 0.162 µl ind–1 h–1. The mean CR using the thymidine-labelled natural bacteria and a 10 min feeding time was 0.179 µl ind–1 h–1. Thus, the CR based on the microsphere method was twice as high as for the other two methods. 相似文献
196.
For products derived from continuous cell lines, regulatory agencies worldwide require that the purification process be validated for its ability to remove or inactivate potential contaminants such as viruses and virus-like particles. New guidance suggests a requirement for statistical evaluation of these studies but the industry has yet to develop such standards. The task of estimating excess capacity is also complicated by variable assays, accumulation of variability in clearance estimates over unit operations, dependence of clearance capacity on operating parameters, and expense of experiments. We propose an experimental strategy to determine the excess clearance capacity of a biopharmaceutical process and to provide statistical estimation of excess capacity in an efficient way. Clearance estimates and their variances are calculated for each orthogonal unit operation and estimates are combined to form an interval estimate of overall process clearance capacity. Poisson regression is suggested as an efficient technique for data analysis of clearance studies. We believe that this approach should meet regulatory guidelines in a cost effective way, while clarifying the roles of qualitative and quantitative components in setting requirements. 相似文献
197.
198.
Freshwater, male Atlantic stingrays Dasyatis sabina , from Lake Jesup, Florida, U·S·A·, excreted a dilute urine similar in composition to freshwater teleosts and lampreys with the exception that urea was the primary osmolyte. Urine flow rate was 2·5 to 10 fold higher than that reported for freshwater teleosts resulting in high free-water clearance. Mass-specific free-water clearance values from euryhaline elasmobranchs inhabiting freshwater environments greatly exceed those for freshwater teleosts and are nearly equivalent to those of freshwater lampreys. 相似文献
199.
200.
Clearance rates of Limnoperna fortunei (Bivalvia) were investigated in laboratory experiments using monocultures of the alga Chlorella vulgaris. Experimental conditions included two mollusc sizes (15 and 23 mm), and three water temperatures (15, 20 and 25 °C) covering the normal seasonal range in the lower Paraná river and Río de la Plata estuary. Filtration rates obtained were, for the larger mussels: 9.9, 13.1 and 17.7 ml mg tissue dry weight–1 h–1 at 15, 20 and 25 °C, respectively; and for the smaller ones: 17.7, 20.8 and 29.5 ml mg–1 h–1. Differences between sizes and between temperatures (except 15 vs. 20 °C) were statistically significant. In absolute terms larger animals have higher clearance rates, but as a function of body mass smaller individuals feed more actively. Within the range of experimental values used, filtration rates were positively associated with water temperature. These clearance rates (125–350 ml individual–1 h–1) are among the highest reported for suspension feeding bivalves, including the invasive species Dreissena polymorpha, D. bugensis and Corbicula fluminea. High filtration rates, associated with the very high densities of this mollusc in the Paraná watershed (up to over 200,000 ind m–2) suggest that its environmental impact may be swiftly changing ecological conditions in the areas colonized. 相似文献