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- Faber, Catherine, et al.
(författare)
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Three-dimensional structure of a human Fab with high affinity for tetanus toxoid
- 1998
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Ingår i: Immunotechnology. - 1380-2933. ; 3:4, s. 253-270
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Tidskriftsartikel (refereegranskat)abstract
- Background: The wide range of antibody specificity and affinity results from the differing shapes and chemical compositions of their binding sites. These shapes range from discrete grooves in antibodies elicited by linear oligomers of nucleotides and carbohydrates to shallow depressions or flat surfaces for accommodation of proteins: peptides and large organic compounds. Objectives: To determine the Fab structure of a high-affinity human antitoxin antibody. To explore structural features which enable the antibody to bind to intact tetanus toxoid, peptides derived from the sequence of the natural immunogen and antigenic mimics identified by combinatorial chemistry. To explain why this Fab shows a remarkable tendency to produce crystals consistently diffracting to d spacings of 1.7-1.8 Å. To use this information to engineer a strong tendency to crystallize into the design of other Fabs. Study design: The protein was crystallized in hanging or sitting drops by a microseeding technique in polyethylene glycol (PEG) 8000. Crystals were subjected to X-ray analysis and the three-dimensional structure of the Fab was determined by the molecular replacement method. Interactive computer graphics were employed to fit models to electron density maps, survey the structure in multiple views and discover the crystal packing motif of the protein. Results: Exceptionally large single crystals of this protein have been obtained, one measuring 5 x 3 x 2 mm (l x w x d). The latter was cut into six irregular pieces, each retaining the features of the original in diffracting to high resolution (1.8 Å) with little decay in the X-ray beam. In an individual Fab, the active site is relatively flat and it seems likely that the protein antigen and derivative peptides are tightly held on the outer surface without significant penetration into the interior. There is no free space to accommodate even a dipeptide between V(H) and V(L). One of the unique features of the B7-15A2 Fab is a large aliphatic ridge dominating the center of the active site. The CDR3 of the H chain contributes significantly to this ridge, as well as to adjoining regions projected to be important for the docking of the antigen. Both the ease of crystallization and the favorable diffraction properties are mainly attributable to the tight packing of the protein molecules in the crystal lattice. Discussion: The B7-15A2 active site provides a stable and well defined platform for high affinity docking of proteins, peptides and their mimotopes. The advantages for future developments are suggested by the analysis of the crystal properties. It should be possible to incorporate the features promoting crystallization, close packing and resistance to radiation damage into engineered human antibodies without altering the desired specificities and affinities of their active sites.
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2. |
- Feng, Bo, et al.
(författare)
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Mechanisms of N2O Formation from Char Combustion
- 1996
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Ingår i: Energy & Fuels. - 1520-5029 .- 0887-0624. ; 10:1, s. 203-208
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Tidskriftsartikel (refereegranskat)abstract
- This paper describes an experimental study on N2O and NOx emission from char combustion. Experiments have been carried out in a fixed bed reactor in the temperature range of 973−1323 K. Oxygen concentrations for combustion, temperature, and char type have been found to have strong effects on both N2O and NOx emissions. With an increasing temperature, N2O reaches a peak at 1073 K and then decreases with temperature. At the same time NOx remains nearly the same. When oxygen concentration increases, the concentration of N2O increases and NOx first increases and then decreases. The char with higher nitrogen content emits more N2O, indicating that nitrogen oxides come from char-N. When NO is included in the inlet gases, much more N2O is produced from char combustion. This suggests that the reaction of NO + O2 + char is the main pathway for N2O formation. Another pathway, i.e., the homogeneous oxidation of HCN from the gasfication of char, seems to be important as well.
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3. |
- Feng, Bo, et al.
(författare)
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Nitrogen oxides emission from a circulating fluidized bed combustor
- 1998
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Ingår i: International Journal of Energy Research. - 1099-114X .- 0363-907X. ; 20:11, s. 1015-1025
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Tidskriftsartikel (refereegranskat)abstract
- Experiments were carried out in a pilot-scale circulating fluidized bed (CFB) coal combustor to investigate the mechanism of N,O formation, nitrogen oxides (including NO, and N,O) emission and the effect of temperature, excess air ratio, recirculation ratio, etc. The concentrations of nitrous oxide and nitric oxide were measured along the height of the CFB furnace. N,O concentration increased with height, and in the exit of the combustor N,O reached the highest level. NO,, however, decreased with height, showing the inverse trend compared with N,O. The N,O emission decreased sharply with the rise of temperature at the bottom of the combustor; at the same time, the NO, concentration increased.
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