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P H Ehrlich W R Moyle Z A Moustafa 《Journal of immunology (Baltimore, Md. : 1950)》1983,131(4):1906-1912
We reported previously that mixtures of some monoclonal antibodies directed against the glycoprotein hormone human chorionic gonadotropin (hCG) had a higher affinity for the antigen than either monoclonal antibody separately. The synergistic interaction could no longer be detected when one of the antibodies was replaced with its F(ab) fragment. This cooperative interaction has now been further characterized. One-half of 10 possible pairs prepared from five IgG1 monoclonal antibodies against hCG result in a synergistic interaction. The addition of an IgG2b monoclonal antibody to one of the IgG1 monoclonal antibodies also induces a cooperative interaction, which shows that the effect is not subclass restricted. Cooperative interactions between antibodies are also not restricted to solution conditions; adsorption of one antibody to a solid support appears to increase the cooperative effect. Indeed, one pair of antibodies that failed to bind hCG synergistically in solution did so when one antibody was bound to a solid surface. The liquid phase antibody also has an effect on the specificity of the solid phase antibody. The sensitivity of the solid phase assay system has enabled us to develop a rapid method of determining if two monoclonal antibodies can bind to an antigen simultaneously. A quantitative theoretical model has been devised that successfully predicts the cooperative behavior observed between antibodies and should be useful in devising conditions that result in sensitive solid phase radioimmunoassays. 相似文献
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The work-to-fracture of canine femoral bone has been measured using the technique of Tattersall and Tappin (1966). The work required to fracture a specimen in three point bending by slow crack propagation through a triangular cross section is obtained from the load-deformation curve. The area of the resulting fracture surface is measured by macrophotographic techniques, and the work-to-fracture is calculated as work per unit area. The values of fracture “toughness” measured in this way ranged from 5.36 × 103 J/m2 to 1.55 × 104 J/m2 in the samples tested with a mean of 9.03 × 103 J/m2 and a standard deviation of 3.27 × 103 J/m2. The work-to-fracture was found to vary with transverse variation in location in the femoral shaft. Scanning electron microscope photographs of the fracture surfaces indicate that the nature of the failure is similar to that of fiber reinforced composite materials. Samples which failed by catastrophic crack propagation were characterized by smooth fracture surfaces and had larger osteons than those which failed by slow crack propagation. 相似文献
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Oligonucleotide-directed mutagenesis was used to complete a collection of mutations in the -35 and -10 hexamers of the ant promoter of Salmonella phage P22. The effects of all 36 single-base-pair substitutions on promoter strength in vivo were measured in strains carrying the mutant promoters fused to an ant-lacZ gene on a single-copy prophage. The results of these assays show that certain consensus base pairs are more important than others; in general, the least-critical positions are among the most poorly conserved. Some mutations within the hexamers have smaller effects on promoter strength than certain mutations outside the hexamers in this and other promoters. Several different patterns of base pair preferences are observed. These hierarchies of base pair preferences correlate well (but not perfectly) with the hierarchies defined by the frequency distribution of base pairs at each position among wild-type promoters. The hierarchies observed in the ant promoter also agree well with most of the available information on base pair preferences in other promoters. 相似文献
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W R Moyle M M Matzuk R K Campbell E Cogliani D M Dean-Emig A Krichevsky R W Barnett I Boime 《The Journal of biological chemistry》1990,265(15):8511-8518
The glycoprotein hormones are a family of conserved heterodimeric proteins which share a common alpha subunit but differ in their hormone-specific beta subunits. We used chimeras of human chorionic gonadotropin (hCG) and luteinizing hormone (hLH) beta subunits to identify residues which enable monoclonal antibodies (mAb) to distinguish the two hormones. The LH beta-CG beta chimeras appeared to fold similar to hCG beta, since they combined with hCG alpha and, depending on their sequences, were recognized by hCG-selective mAbs. Amino acid residues Arg8-Arg10,Gly47-Ala51, and Gln89-Leu92 form a major epitope region and appear to be adjacent to each other on the surface of hCG beta. Gly47-Ala51 and Gln89-Leu92 are recognized by dimer-specific mAbs while Arg8-Arg10 is recognized by mAbs which have highest affinity for the free beta subunit. These observations suggest that the conformation of this region of the beta subunit changes when the alpha and beta subunits combine. Residues which are C-terminal of Asp112 form a second epitope domain. mAbs to the third domain distinguish hCG beta and hLH beta by the presence of Asn77 in hCG beta and can be detected after hCG binds to receptors. These findings were used to develop a model of hCG beta which predicts the locations of these residues and their positions relative to the alpha subunit and receptor interfaces. 相似文献
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Synopsis The life history characteristics of tule perch were compared within and among populations in three drainages with substantially different environmental conditions. Within populations there were, in general, substantial increases in brood size, size of young, brood weight, and gonadal-somatic index (GSI) with age and with size of female. In at least one population there was a trade-off between the number and size of young produced. However, in a lake where individual growth was very slow, brood weight, GSI, and other life history characters showed decreasing trends with age. Among populations in isolated drainages, female length at first reproduction and longevity varied directly and mean brood size varied inversely with environmental predictability. Morphological and geological evidence and some comparative litter characteristics suggest that life history differences among populations in isolated drainages are in part genetic. 相似文献
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A rat ovarian cDNA library was constructed and screened by differential colony hybridization to detect cDNA clones specific for mRNA induced by follicle-stimulating hormone (FSH). The cDNA clone which demonstrated the greatest degree of induction contained a 766-bp insert which was characterized and sequenced. We conclude that this cDNA is specific for the rat gene coding for cholesterol side-chain cleavage enzyme (P-450scc) by virtue of nucleotide sequence homology to the bovine and human P-450scc cDNA sequences. Southern blotting of rat genomic DNA suggests the presence of a single P-450scc gene. Northern blot analysis indicates that P-450scc mRNA is present in steroidogenic tissues (ovary, adrenal, testis), but not in brain, kidney, liver, lung, or heart. The rat P-450scc mRNA is induced by FSH or pregnant mare's serum gonadotropin in ovaries of estrogen-treated immature rats in vivo. In cultured granulosa cells, estradiol treatment alone did not increase P-450scc mRNA levels, but in combination with FSH or 8-Br-cAMP resulted in three- to four-fold increase in this mRNA. 相似文献