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41.
Regarding biological products, increasing awareness of potential side effects have placed great importance not only at protein
purity regarding other proteins but on the removal of biologicals such as DNA and especially virus the importance of which
may not be known. Monoclonal antibodies (Mab) have come to be an important class of molecules obtained from hybridoma cells,
i.e., nonrecombinant cells in culture. It has been noted during the last years, that with rare exceptions hybridoma cell lines
contain retrovirus like particles. The infectious nature of the EM-visible particles has been tested for, however, in most
cases not been substantiated. In order to bring these valuable biological reagents, Mab's, to good use in man for imaging
or therapy, the remaining concern about a potential retroviral infection has to be reduced to an acceptable minimum. We describe
experimental approaches for the validation of chromatographic and ultrafiltration steps used in the production of monoclonal
antibodies to remove and inactivate murine retrovirus.
Present day biotechnological manufacturing processes have been devised incorporating a number of strategic preventive measures
that have found wide spread acceptance. They permit to answer the question: how can a potentially harmful infection by an
unknown virus be excluded.
Knowledge of the efficacy of purification steps to clear infectious model virus is fundamental to devise biotechnological
manufacturing processes yielding a purified antibody for use in man. 相似文献
42.
Tracey C. Bourner Enrique Vargas‐Osuna Trevor Williams Candido Santiago‐Alvarez Jenny S. Cory 《Biocontrol Science and Technology》1992,2(4):315-326
Agrotis segetum nuclear polyhedrosis virus (AsNPV) and granulosis virus (AsGV), propagated in laboratory cultures of A. segetum in England and A. ipsilon in Spain, respectively, were applied to plots of maize plants at the one‐ to four‐leaf stage of growth. Plots were arranged in a 6 x 6 Latin square design and infested with second‐instar A. segetum larvae (the common cutworm). Each virus was applied in separate treatments by two application methods; as an aqueous spray containing 0.1% Agral as a wetting agent, and as a bran bait. The NPV was applied at a rate of 4 X 1012 polyhedra/ha, and the GV at 4 X 1013 granules/ha. Soil and plants were sampled for larvae on three occasions following virus treatment: 24 h, 4 days and 11 days. The larvae were reared on diet in the laboratory, until death or pupation, to examine the rate and level of viral infection. Infection data showed 87.5% and 91% NPV infection and 12.5% and 55% GV infection in spray and bait treatments, respectively, in larvae sampled 24 h after treatment. In larvae sampled 4 days after treatment, the results were 78% and 100% NPV infection, and 13% and 6% GV infection. A total of only six larvae were retrieved on day 11. In both treatments larvae infected with AsNPV died significantly more rapidly and at an earlier instar than those infected with AsGV, indicating that AsNPV appears to have better potential as a control agent for A. segetum. 相似文献
43.
The potato has tremendous potential as a transgenic crop and is a good model system by which to analyse metabolic regulation
and gene expression. The potato’s difficult genetics, but ease of genetic transformation and its clonal means of propagation
make it ideal for applied agricultural molecular genetics. Thus, the next 4 years promise to put the potato (with a diversity
of transgenic constructs expressed) in the limelight as many of the first transgenic agricultural products enter the marketplace. 相似文献
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47.
B. N. Manjula K. M. Khandke T. Fairwell W. A. Relf K. S. Sriprakash 《Journal of Protein Chemistry》1991,10(4):369-384
Streptococcal M protein, a dimeric alpha helical coiled-coil molecule, is an antigenically variable virulence factor on the surface of the bacteria. Our recent conformational analysis of the complete sequence of the M6 protein led us to propose a basic model for the M protein consisting of an extended central coiled-coil rod domain flanked by a variable N-terminal and a conserved C-terminal end domains. The central coiled-coil rod domain of M protein, which constitutes the major part of the M molecule, is made up of repeating heptads of the generalized sequence a-b-c-d-e-f-g, wherein a and d are predominantly apolar residues. Based on the differences in the heptad pattern of apolar residues and internal sequence homology, the central coiled-coil rod domain of M protein could be further divided into three subdomains I, II, and III. The streptococcal sequelae rheumatic fever (RF) and acute glomerulonephritis (AGN) have been known to be associated with distinct serotypes. Consistent with this, we observed that the AGN associated M49 protein exhibits a heptad motif that is distinct from the RF associated M5 and M6 proteins. Asn and Leu predominated in the a and d positions, respectively, in subdomain I of the M5 and M6 proteins, whereas apolar residues predominated in both these positions in the M49 protein. To establish whether the heptad motif of M49 is unique to this protein, or is a general characteristic of nephritis-associated serotypes, the amino acid sequence of M57, another nephritis-associated serotype, has now been examined. The gene encoding M57 was amplified by PCR, cloned into pUC19 vector, and sequenced. The C-terminal half of M57 is highly homologous to other M proteins (conserved region). In contrast, its N-terminal half (variable region) revealed no significant homology with any of the M proteins. Heptad periodicity analysis of the M57 sequence revealed that the basic design principles, consisting of distinct domains observed in the M6 protein, are also conserved in the M57 molecule. However, the heptad motif within the coiled-coil subdomain I of M57 was distinct from M5 and M6 but similar to M49. Similar analyses of the heptad characteristics within the reported sequences of M1, M12, and M24 proteins further confirmed the conservation of the overall architectural design of sequentially distinct M proteins. Furthermore, the heptad motif within subdomain I of the AGN-associated serotypes M1 and M12 was similar to M49 and M57, whereas that of the RF associated M24 was similar to the M5 and M6 proteins. These results clearly demonstrate a correlation between the heptad motifs within the distal coiled-coil subdomain of the M proteins from different streptococcal serotypes and their epidemiological association with the sequelae AGN and RF. 相似文献
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In field experiments, larvae of codling moth Cydia pomonella (L.) rarely acquired granulosis virus on hatching from the egg, but picked up most later, on the tree surface. Deposits of virus sprayed in 1.0% w/v skimmed milk did not affect neonate larval behaviour. Larvae died, usually in the first instar, after entering treated fruit, but they frequently entered via the calyx or near the base of the stalk or through cracks in the skin, where little feeding damage by first-and sometimes second-instar larvae was seen.
Résumé En verger, la pulvérisation d'oeufs de carpocapse avec du virus de la granulose en suspension dans l'eau (additionnée de lait écrémé dilué à 1%) n'a pas modifié la survie des chenilles avant pénétration dans le fruit; par contre la pulvérisation des arbres a provoqué une forte mortalité. Bien que des chenilles consommant des poils et la surface des feuilles aient été observées avant leur pénétration dans le fruit, ce qui aurait pu provoquer leur contamination par le virus, il semble que la contamination létale provienne des fruits seuls.La présence de produit n'a modifié ni le comportement larvaire, ni le taux de pénétration dans les fruits; la mortalité y a lieu ensuite, généralement au premier stade. Dans 74 à 78% des cas, les chenilles ont pénétré dans le fruit par le calice ou près de la base du pédoncule — aucun dégât provenant de larves du premier stade n'y était visible, de même que dans le calice pour les larves du deuxième stade. Par contre, toute pénétration par la surface du fruit était repérable dès le premier stade. Il est possible que la répartition des lieux de pénétration dans le fruit influe sur la létalité due au virus et explique les variations d'efficacité observées en verger. Un système de classification des dégâts, provoqués lors de la pénétration dans le fruit, de chenilles du premier au troisième stade est proposé pour évaluer l'efficacité des essais en verger.相似文献