全文获取类型
收费全文 | 184篇 |
免费 | 11篇 |
出版年
2022年 | 4篇 |
2021年 | 3篇 |
2019年 | 3篇 |
2018年 | 4篇 |
2017年 | 1篇 |
2016年 | 2篇 |
2015年 | 3篇 |
2014年 | 3篇 |
2013年 | 5篇 |
2012年 | 6篇 |
2011年 | 6篇 |
2010年 | 3篇 |
2009年 | 7篇 |
2008年 | 6篇 |
2007年 | 9篇 |
2006年 | 3篇 |
2005年 | 8篇 |
2004年 | 4篇 |
2003年 | 10篇 |
2002年 | 5篇 |
2001年 | 9篇 |
2000年 | 2篇 |
1999年 | 2篇 |
1998年 | 4篇 |
1997年 | 4篇 |
1996年 | 1篇 |
1995年 | 3篇 |
1994年 | 2篇 |
1993年 | 4篇 |
1992年 | 5篇 |
1991年 | 8篇 |
1990年 | 2篇 |
1989年 | 5篇 |
1988年 | 7篇 |
1987年 | 5篇 |
1986年 | 1篇 |
1985年 | 3篇 |
1984年 | 2篇 |
1983年 | 4篇 |
1982年 | 3篇 |
1981年 | 3篇 |
1980年 | 1篇 |
1978年 | 3篇 |
1977年 | 3篇 |
1976年 | 2篇 |
1974年 | 3篇 |
1973年 | 3篇 |
1972年 | 4篇 |
1971年 | 2篇 |
排序方式: 共有195条查询结果,搜索用时 78 毫秒
101.
A semi-synthetic repertoire of intrinsically stable antibody fragments derived from a single-framework scaffold. 总被引:4,自引:0,他引:4
A Desiderio R Franconi M Lopez M E Villani F Viti R Chiaraluce V Consalvi D Neri E Benvenuto 《Journal of molecular biology》2001,310(3):603-615
We report the design, construction and use of an antibody bacteriophage display library built on the scaffold of a single-chain variable fragment (scFv) previously proven to be functionally expressed in the reducing environment of both bacterial and plant cytoplasm and endowed with intrinsic high thermodynamic stability. Four amino acid residues of the third hypervariable loop (CDR3) of both VH and VL were combinatorially mutated, generating a repertoire of approximately 5x10(7) independent scFvs, cloned in a phagemid vector. The ability of the antibody phage library to yield specific binders was tested by biopanning against several antigens. Successful selection of fully active scFvs was obtained, confirming the notion that combinatorial mutagenesis of few amino acid residues centrally located in the antigen-binding site is sufficient to provide binding specificities against virtually any target. High yields of both soluble and phage antibodies were obtained in Escherichia coli. Maintenance of the cognate scFv antibody stability in the newly selected scFv fragments was demonstrated by guanidinium chloride denaturation/renaturation studies and by soluble antibody expression in the bacterial cytoplasm. The antibody library described here allows the isolation of new stable binding specificities, potentially exploitable as immunochemical reagents for intracellular applications. 相似文献
102.
Plant pharming of a full-sized, tumour-targeting antibody using different expression strategies 总被引:1,自引:0,他引:1
Villani ME Morgun B Brunetti P Marusic C Lombardi R Pisoni I Bacci C Desiderio A Benvenuto E Donini M 《Plant biotechnology journal》2009,7(1):59-72
The aims of this work were to obtain a human antibody against the tumour-associated antigen tenascin-C (TNC) and to compare the yield and quality of plant-produced antibody in either stable transgenics or using a transient expression system. To this end, the characterization of a full-sized human immunoglobulin G (IgG) [monoclonal antibody H10 (mAb H10)], derived from a selected single-chain variable fragment (scFv) and produced in plants, is presented. The human mAb gene was engineered for plant expression, and Nicotiana tabacum transgenic lines expressing both heavy (HC) and light (LC) chain were obtained and evaluated for antibody expression levels, in vivo assembly and functionality. Affinity-purified H10 from transgenics (yield, 0.6–1.1 mg/kg fresh weight) revealed that more than 90% of HC was specifically degraded, leading to the formation of functional antigen-binding fragments (Fab). Consequently, H10 was transiently expressed in Nicotiana benthamiana plants through an Agrobacterium -mediated gene-transfer system. Moreover, the use of the p19 silencing suppressor gene from artichoke mottled crinkle virus raised antibody expression levels by an order of magnitude (yields of purified H10, 50–100 mg/kg fresh weight). Approximately 75% of purified protein consisted of full-sized antibody functionally binding to TNC ( K D = 14 n m ), and immunohistochemical analysis on tumour tissues revealed specific accumulation around tumour blood vessels. The data indicate that the purification yields of mAb H10, using a transient expression system boosted by the p19 silencing suppressor, are exceptionally high when compared with the results reported previously, providing a technique for the over-expression of anticancer mAbs by a rapid, cost-effective, molecular farming approach. 相似文献
103.
104.
TMS (trimethylsilyl), MO-TMS (methyl oxime-TMS), and EO-TMS (ethyl oxime-TMS) derivatives of several prostaglandins (A, B1, B2, E1, 8-iso-E1, E2 and 8-iso-E2) were prepared and their gas chromatographic properties examined on a moderately polar (OV-17) and a relatively non-polar (SE-30) stationary phase. Combined gas chromatography-mass spectrometry (GC-MS) using an LKB 9000 instrument was used to identify the different derivatives. Although the TMS derivatives are more easily prepared, the TMS derivatives of the PgE series are thermally somewhat unstable. Thus, MO-TMS and EO-TMS derivatives which exhibit more regular retention increments are more useful for analytical work. The EO-TMS derivatives may be useful in determining mass spectral fragmentation modes of the prostaglandin derivatives. 相似文献
105.
106.
One patient's lumbar CSF sample out of 54 studied for lower back pain had an atypical metabolic profile of opioid receptoractivity. To test our hypothesis that neuropeptides play a role in human lower back pain, endogenous opioid receptoractivity was compared between a control and that atypical sample. Two lumbar puncture samples were obtained from those two patients, one before and one after clinical evaluation. Total opioid receptoractivity was measured in each sample before HPLC separation, and opioid receptoractivity was measured in each fraction after HPLC separation. The latter data represent a metabolic profile of opioid receptoractivity in the human lumbar CSF. Those data demonstrate that the atypical patient had opioid receptoractivity measurements (total and profile) differing in a qualitative and quantitative sense from the other 53 patients studied. CSF opioid receptoractivity reflects the metabolism of opioid neuropeptidergic systems in the physiology in those patients, and opioid peptides may play a role in lower back pain. 相似文献
107.
Kumaran Kandasamy Sujatha S Mohan Rajesh Raju Shivakumar Keerthikumar Ghantasala S Sameer Kumar Abhilash K Venugopal Deepthi Telikicherla Daniel J Navarro Suresh Mathivanan Christian Pecquet Sashi Kanth Gollapudi Sudhir Gopal Tattikota Shyam Mohan Hariprasad Padhukasahasram Yashwanth Subbannayya Renu Goel Harrys KC Jacob Jun Zhong Raja Sekhar Vishalakshi Nanjappa Lavanya Balakrishnan Roopashree Subbaiah YL Ramachandra Abdul B Rahiman Keshava TS Prasad Jian-Xin Lin Jon CD Houtman Stephen Desiderio Jean-Christophe Renauld Stefan N Constantinescu Osamu Ohara Toshio Hirano Masato Kubo Sujay Singh Purvesh Khatri Sorin Draghici Gary D Bader Chris Sander Warren J Leonard Akhilesh Pandey 《Genome biology》2010,11(1):1-9
108.
Emanuela Callone Marco Ischia Angiola Forleo Roberto Paolesse 《Inorganica chimica acta》2008,361(1):79-85
SnO2 and SnO2 + Co-porphyrin solids were prepared from SnCl4 in propanol and hydrolyzed to sol. Thermal behavior of samples obtained at 110 °C was studied in the 20-600 °C interval by thermal analysis coupled with mass spectrometry for identification of released species. The original samples maintain residual Sn-OR, Sn-OH and Sn-Cl groups up to 350 °C. The sample doped with 1% Co-porphyrin differs for a significant presence of residual Sn-Cl species, accounting for SnCl4 release in the 300-340 °C range.119Sn solid state NMR analysis reveals disordered SnO2 species in the sample heated at 250 °C and non-uniform SnO6 units in the SnO2 + Co-porphyrin sample at 110 °C, due to persistence of Sn-OR and Sn-OH groups. This complexity is lost at 250 °C. X-ray diffraction analysis confirms all these data. The sensing efficiency of these materials versus alcohols is ascribed to the presence of an open, incomplete SnO2 structure, which is more pronounced in the Co-porphyrin-doped sample. 相似文献
109.
110.