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排序方式: 共有187条查询结果,搜索用时 15 毫秒
81.
V. V. Argentova T. K. Aliev V. A. Toporova V. S. Rybchenko D. A. Dolgikh M. P. Kirpichnikov 《Moscow University Biological Sciences Bulletin》2017,72(2):63-68
Production and application of therapeutic monoclonal antibodies are second only to vaccines in the world pharmaceutical market. The most common therapeutic antibodies are monoclonal antibodies (mAbs) of the IgG isotype that are produced in eukaryotic CHO cells. In recent years, there has been a considerable interest in developing treatment medications based on IgA antibodies, which can have a wide range of effector functions on human mucous membranes. To study the expression level of immunoglobulin A (IgA) in mammal cells, we designed a set of bipromoter (CMV and EF1α) vectors. The vectors contain gene fragments that encode the heavy chain variable domain (VH) and the light chain variable domain (VL) of the human monoclonal antibody FI6v3 against the hemagglutinin of influenza virus A. They also contain gene fragments that encode the light chain (kappa type) constant domain and the heavy chain constant domain of the human antibody IgA1. The expression vectors differed in the orientation of the promoters and the presence or absence of introns. Two variants of the full-length light and heavy chains were cloned into a eukaryotic expression vector in head-to-head and head-to-tail orientations. The resulting plasmids were transfected into CHO-DG44 and HEK-293T cells. The antibody expression level for the stable transfection of CHO-DG44 and HEK-293T cell cultures was determined by ELISA. The results of the experiments showed that the expression of FI6v3-IgA1 antibodies significantly increased when eukaryotic cells were transfected with the plasmid pBiPr-ABIgA1FI6-Iht in which the heavy chain of IgA1 contains introns and the promoters are arranged head-to-tail. 相似文献
82.
Perry G Nunomura A Raina AK Aliev G Siedlak SL Harris PL Casadesus G Petersen RB Bligh-Glover W Balraj E Petot GJ Smith MA 《Neurochemical research》2003,28(10):1549-1552
Most studies of Alzheimer's disease (AD) have focused on a single precipitating alteration as the etiological event rather than global changes closely linked to aging. Recent evidence suggests that the most significant of these global changes are metabolic. Here we present data indicating that metabolic rate, nutrition, and neuronal size are all early indicators of AD. Understanding the cellular and molecular basis for these changes may open a new dimension to understanding AD. 相似文献
83.
Mitochondria as a primary target for vascular hypoperfusion and oxidative stress in Alzheimer's disease 总被引:3,自引:0,他引:3
It has been widely accepted that vascular hypoperfusion induces oxidative stress and the outcome of this misbalance is brain energy failure. This abnormality leads to neuronal death which manifests as cognitive impairment and the development of brain pathology as in Alzheimer's disease (AD). It has been demonstrated that the AD brain is characterized by impairments in energy metabolism. We theorize that hypoperfusion induced mitochondrial failure plays a key role in the generation of reactive oxygen species, resulting in oxidative damage to brain cellular compartments, especially in the vascular endothelium and in selective population of neurons with high metabolic activity in the AD brain. All of these abnormalities have been found to occur before classic AD pathology inducing neuronal degeneration and amyloid deposition during the progression of AD. Therefore, expanding investigations into both the mechanisms behind amyloid beta (Abeta) deposition and the possible accelerating effects of environmental factors such as chronic hypoxia/reperfusion may open a new avenue for effective treatments of AD. Future studies examining the importance of mitochondrial pathobiology in brain cellular compartments provide insight not only into the better understanding of the neurodegenerative and/or cerebrovascular disease but also provide targets for treating these conditions. 相似文献
84.
Allakhverdiev SI Hayashi H Nishiyama Y Ivanov AG Aliev JA Klimov VV Murata N Carpentier R 《Journal of plant physiology》2003,160(1):41-49
The presence of 1.0 mol/L glycinebetaine during isolation of D1/D2/Cytb559 reaction centre (RC) complexes from photosystem II (PSII) membrane fragments preserved the photochemical activity, monitored as the light-induced reduction of pheophytin and electron transport from diphenylcarbazide to 2.6-dichlorophenol-indophenol.-Glycinebetaine also protected the D1/D2/Cytb559 complexes against strong light-induced damage to the photochemical reactions and the irreversible bleaching of beta-carotene and chlorophyll. The presence of glycinebetaine also enhanced thermotolerance of the D1/D2/Cytb559 complexes isolated in the presence of 1.0 mol/L betaine with an increase in the temperature for 50% inactivation from 29 degrees C to 35 degrees C. The results indicate an increased supramolecular structural stability in the presence of glycinebetaine. 相似文献
85.
Shingarova LN Tikunova NV Iun TE Chepurnov AA Aliev TK Batanova TA Boldyreva EF Nekrasova OV Toporova VA Panina AA Kirpichnikov MP Sandakhchiev LS 《Bioorganicheskaia khimiia》2007,33(6):598-605
A full-size human antibody to Ebola virus was constructed by joining genes encoding the constant domains of the heavy and light chains of human immunoglobulin with the corresponding DNA fragments encoding variable domains of the single-chain antibody 4D1 specific to Ebola virus, which was chosen from a combinatorial phage display library of single-strand human antibodies. Two expression plasmids. pCH1 and pCL1, containing the artificial genes encoding the light and heavy chains of human immunoglobulin, respectively, were constructed. Their cotransfection into the human embryonic kidney cell line HEK293T provided the production of a full-size recombinant human antibody. The affinity constant for the antibody was estimated by solid-phase enzyme-linked immunoassay to be 7.7 x 10(7) +/- 1.5 x 10(7) M(-1). Like the parent single-chain antibody 4DI, the resulting antibody bound the nucleoprotein of Ebola virus and did not interact with the proteins of Marburg virus. 相似文献
86.
A.?A.?PaninaEmail author I.?G.?Dement’yeva T.?K.?Aliev V.?A.?Toporova M.?N.?Bokov L.?P.?Pozdnyakova O.?B.?Shemchukova D.?A.?Dolgikh P.?G.?Sveshnikov M.?P.?Kirpichnikov 《Russian Journal of Bioorganic Chemistry》2017,43(4):409-420
Previously, we have determined the nucleotide and amino acid sequences of the variable domains of three mouse monoclonal antibodies specific to the individual epitopes of the Ebola virus glycoprotein: GPE118 (IgG), GPE325 (IgM) and GPE534 (IgG) [1]. In the present paper, chimeric Fab fragments of Fab118, Fab325, and Fab534 antibodies were obtained based on the variable domains of murine antibodies by attaching CH1 and CL constant regions of human kappa-IgG1 to them. The recombinant chimeric Fab fragments were synthesized in the heterologous expression system Escherichia coli, isolated and purified using metal chelate affinity chromatography. The immunochemical properties of the obtained Fab fragments were studied by immunoblotting techniques as well as indirect and competitive ELISA using recombinant Ebola virus proteins: EBOV rGPdTM (recombinant glycoprotein of Ebola hemorrhagic fever virus without the transmembrane domain), NP (nucleoprotein) and VP40 (structural protein). The identity of recombinant chimeric Fab fragments, as well as their specificity to the recombinant glycoprotein of Ebola hemorrhagic fever virus (EBOV GP) was proved. The results of indirect ELISA evidence the absence of immunological cross-reactivity to NP and VP40 proteins of Ebola virus. The dissociation constants of the antigen-antibody complex K d equal to 5.0, 1.0 and 1.0 nM for Fab118, Fab325 and Fab534, respectively, were determined; they indicate high affinity of the obtained experimental samples to EBOV GP. The epitope specificity of Fab fragments was studied using a panel of commercial neutralizing antibodies. It was found that all studied antibodies to EBOV GP are targeted to different epitopes, while the epitopes of the recombinant chimeric Fab fragments and original murine monoclonal antibodies (mAbs) coincide. All the obtained and studied mAbs to EBOV GP are specific to epitopes that coincide or overlap the epitopes of three commercial neutralizing mAbs to Ebola virus: epitopes Fab118 and Fab325 overlap the epitope of the known commercial mAb h13F6; Fab325 epitope also overlaps mAb c6D8 epitope; Fab534 epitope is located near mAb KZ52 conformational epitope, in the formation of which amino acid residues of GP1 and GP2 domains of EBOV GP are involved. 相似文献
87.
88.
Babadzhanova M. P. Babadzhanova M. A. Aliev K. A. 《Russian Journal of Plant Physiology》2002,49(5):592-597
Free and membrane-bound forms of Calvin-cycle multienzyme complexes with a mol wt of 520 ± 20 kD and 640 ± 25 kD, respectively, were isolated from the cotton (Gossypium hirsutum L.) leaves. Both complexes exhibited the following enzymatic activities: ribose phosphate isomerase, phosphoribulokinase, ribulose bisphosphate carboxylase (Rubisco), phosphoglycerate kinase, and glyceraldehyde phosphate dehydrogenase. The activities of the membrane-bound multienzyme complex were significantly higher than the activities of the free complex. This difference was especially pronounced in the case of carboxylase activity. An increase in the enzymatic activity of membrane-bound multienzyme complex in comparison with the free complex is presumably due to the different number of their constituent parts. Another possible cause is the membrane-level regulation of the functional activity of the enzymes composing the complex. 相似文献
89.
90.