• Korean Journal of Microbiology
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• v.39 no.4
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• pp.242-247
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• 2003

Murine encephalomyocarditis virus (EMCV) has been used as a surrogate for hepatitis A virus (HAV) for the validation of virus removal and/or inactivation during the manufacturing process of biopharmaceuticals. Recently international regulation for the validation of HAV safety has been reinforced because of the reported cases of HAV transmission to hemophiliac patients who had received ntihemophilic factors prepared from human plasma. The purpose of the present study was to compare the resistance of HAV and EMCV to various viral inactivation processes and then to standardize the HAV validation method. HAV was more resistant than EMCV to pasteurization (60oC heat treatment for 10 hr), low pH incubation (pH 3.9 at 25oC for 14 days), 0.1 M NaOH treatment, and lyophilization. EMCV was completely inactivated to undetectable levels within 2 hr of pasteurization, however, HAV was completely inactivated to undetectable levels after 5 hr treatment. EMCV was completely inactivated to undetectable levels within 15 min of 0.1 M NaOH treatment, however, residual infectivity of HAV still remained even after 120 min of treatment. The log reduction factors achieved during low pH incubation were 1.63 for HAV and 3.84 for EMCV. Also the log reduction factors achieved during a lyophilization process of antihemophilic factor VIII were 1.21 for HAV and 4.57 for EMCV. These results indicate that HAV rather than EMCV should be used for the virus validation study and the validation results obtained using EMCV should be precisely reviewed.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.340-346
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• 2002

The purpose of the present study was to examine the efficacy and mechanism of the PAB (para-amino benzamidine) affinity column chromatography, Viresolve NFP virus filtration, pasteurization (60$\^{C}$ heat treatment for 10 h), and lyophilization steps employed in the manufacture of urokinase from human urine as regards the removal and/or inactivation of the hepatitis A virus (HAV). Samples from the relevant stages of the production process were spiked with HAV and subjected to scale-down processes mimicking the manufacture of urokinase Samples were collected at each step, immediately titrated using a 50% tissue culture infectious dose (TCID$\_$50/), and the virus reduction factors evaluated. PAB chromatography was found to be an effective step for removing HAV with a log reduction factor of 3.24. HAV infectivity was rarely detected in the urokinase fraction, while most of the HAV infectivity was recovered in the unbound and wash fractions. HAV was completely removed during the Viresolve NFP filtration with a log reduction factor of $\geq$ 4.60. Pasteurization was also found to be an effective step in inactivating HAV where the titers were reduced from an initial titer of 7.18 log$\_$10/ TCID$\_$50/ to undetectable levels within 10 h of treatment. The log reduction factor achieved during pasteurization was $\geq$ 4.76. Lyophilization revealed the lowest efficacy for inactivating HAV with a log reduction factor of 1.48. The cumulative log reduction factor was $\geq$ 14.08. Accordingly, these results indicate that the production process for urokinase exhibited a sufficient HAV reducing capacity to achieve a high margin of virus safety.

• Korean Journal of Veterinary Service
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• v.16 no.1
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• pp.20-33
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• 1993

The studies were conducted to determine the effects of preservatives such as sodium chloride, sodium nitrite, sodium benzoate and sorbic acid on the survival of L. monocytogenes with regard to interaction of temperature, heat and pH of the medium. Inactivation of L. monocytogenes Scott A was more predominent by combination of sodium chloride and the other preservatives than sodium chloride alone, and inactivation was more exhilarated at $4^{\circ}C$ than at $35${\circ}C.$ The organism was not inactivated when sodium chloride, sodium nitrite, sodium benzoate and sorbic acid were added to 3%, 100ppm, 0.1, or lower, respectively, but was inactivated in the concentration increased twice. In TSB(tryptic soy broth) at pH 5.0 or lower, the organism did not grow regadless of the kinds of preservatives, and inactivation effect particularly was prominent in the presence of sodium nitrite and sorbic acid. On the other hand, at pH 6.0 or higher L. monocytogenes gradually increased in numbers and the effects of inhibition was higher in the presence of sorbic acid than in the other preservatives. When the preservatives were added to the concentration commonly used, incubation in TSB at $4^{\circ}C$ gradually resulted in growth of the bacterium and the organism rapidly decreased in numbers at $20^{\circ}C\; or\; 35^{\circ}C$ after incubation for 1 week. When L. monocytogenes was inoculated in TSB containing various preservatives and heated at $55^{\circ}C$ for 30minutes, the organism decreased in numbers at all preservatives. Particularly, viability rate of the organism was the lowest as 0.07% in the presence of sorbic acid.

• Journal of Plant Biotechnology
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• v.42 no.1
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• pp.43-48
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• 2015

It is a serious situation that the farmers' income has gradually decreased due to the decline of productivity of fruit trees infected with viroids. It has been known that Persimmon viroid (PVd) and Citrus viroid (CVd) are economically important viroids that can infected persimmon. In this study, the incidence of CVd and PVd in 'Fuyu' persimmon were identified as 41% and 34% in JeollaNam-do, respectively. The collected persimmon samples infected by both PVd and CVd were used for testing efficiency of the viroid inactivation methods. The samples were subjected to single treatment of the heat treatment ($37^{\circ}C$), cold treatment ($4^{\circ}C$), or antiviral agent treatment (Ribavirin), and double treatment of combinations of the three methods. Viroid inactivation efficiency was confirmed through RT-PCR. In the case of the samples subjected to cold treatment for 4 weeks, the viroid inactivation efficiency was most significantly high as 67% against the survival rate of 100%. In addition, in the case of the samples treated for 2 weeks with the antiviral agents and cold treatment, the viroid inactivation rate was similar to that of the cold treatment. In conclusion, the cold treatment showed the highest viroid inactivation efficiency, and this result will provide valuable information for production of viroid-free persimmon.

• Proceedings of the Membrane Society of Korea Conference
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• 1996.10a
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• pp.18-21
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• 1996

Virus and DNA removal in bio-drug manufacturing processes has received a great deal of attention in recent years. Removing of a virus using a membrane process is a promising method, because inactivated virus can be removed from the bio-drug and the process can be used as an additional and security inactivation after the method of general heat-inactivation of the virus in the bio-drug. The FDA and the biopharmaceutical industry have recently announced strict guidelines for impurities of virus and DNA contamination. The regulatory guidelines on residual amounts of DNA in mammalian cell culture products require DNA contamination of less than 100 pg/dose. Therefore, permeation and rejection of DNA through the porous membranes have become important in the application of DNA removal in bio-drug manufacturing using membrane technology. In this study, the permeation of DNA and chromatin through regenerated cellulose hollow fibers that have a mean pore diameter of 15 nm was investigated.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.615-618
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• 2000

The purpose of present study was to examine the efficacy of PAB (para-amino benzamidine) affinity column chromatography, pasteurization ($60^{\circ}C$ heat treatment for 10 h), and lyophilization steps, employed in the manufacture of urokinase from human urine, in the removal and/or inactivation of urine-born viruses. Bovine herpes virus (BHV) and Murine encephalomyocarditis virus (EMCV) were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses and the amount of virus in each fraction was quantified by 50% tissue culture infectious dose ($TCID_{50}$). BHV and EMCV were effectively partitioned from urokinase during PAB chromatography with the log reduction factors of 6.71 and 5.27, respectively. Pasteurization was a robust and effective step in inactivating BHV and EMCV, of which titers were reduced from initial titers of $8.65\;log_{10}\;TCID_{50}$ and $7.81\;log_{10}\;TCID_{50}$, respectively, to undetectable levels within 1 hour of treatment. The log reduction factors achieved during lyophilization were 2.06 for BHV and 4.54 for EMCV. These results indicate that the production process for urokinase has sufficient virus reducing capacity to achieve a high margin of virus safety.

• Biomedical Science Letters
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• v.23 no.3
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• pp.230-237
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• 2017

Changes in the activity of alkaline phosphatase (ALP) isoenzymes and isoforms in human serum have a major diagnostic value, therefore the regulation of ALP activities is a valuable target for therapeutic interventions. To assess the pharmacological activity of retinoids, i.e., all-trans retinoic acid and 13-cis retinoic acid, their tissue-specific inhibitory effect on human serum ALP activity was elucidated by chemical inhibition methods, heat-sensitive inactivation, and wheat-germ lectin precipitation test. Retinoids showed significant inhibition of the total ALP activity in human serum at a concentration of 5 mM. All-trans retinoic acid (5 mM) and 13-cis retinoic acid (5 mM) inhibited ALP activities by up to 12% and 15%, respectively, compared to that by guanidine hydrochloride (200 mM). L-phenylalanine (100 mM) and urea (30 mM) had no further inhibitory effect on ALP activities in human serum pretreated with retinoids (5 mM). Retinoids significantly inhibited ALP activities by up to 20% compared with that of tetramisole (30 mM). The ALP activities in retinoid-pretreated serum remained unchanged after the heat inactivation process. These results suggest that retinoids are inhibitors of the intestinal ALP isoenzyme. Remarkably, retinoids revealed potent inhibitory activities against ALP in wheat-germ lectin precipitant serum, indicating that they also function as inhibitors of the bone ALP isoform. The results show that retinoids inhibit the specific tissue-derived human serum ALP activities, moreover, the inhibitory effect of retinoids against bone ALP activity suggests their clinical utility as monitoring and prevention of metastasis of bone cancer.

• Korean journal of food and cookery science
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• v.14 no.5
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• pp.492-497
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• 1998

The stabilizing effect of fructooligosanharide (FO) on hen＇s egg yolk immunoglobulin (yIgG) by heat and acid was investigated. The heat stability of yIgG at 70∼80$^{\circ}C$ was enhanced in a concentration-dependent manner by adding 0∼50% (w/v) FO to a yIgG solution. Acid-induced inactivation of yIgG was also suppressed in a concentration-dependent relationship by addition of FO. Addition of 50% FO almost completely stabilized yIgG at pH 3. The remarkable stablizing effect of FO on yIgG may enhance the use of yIgG as functional food ingredients.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.713-717
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• 2012

The objective of this study was to evaluate the effect of sodium habituation on thermal resistance of Staphylococcus aureus in various ready-to-heat (RTH) sauces. The strain mixture of S. aureus strains KACC10768, KACC10778, KACC11596, KACC13236 and NCCP10862 was habituated up to 9% of NaCl. The inocula of NaCl-habituated and non-habituated S. aureus were inoculated in 5 g portions of pork cutlet, meat and Carbonara sauces at 7 Log CFU/g, and the samples were vortexed vigorously. The inoculated samples were then exposed to 60 and $70^{\circ}C$ in a water-bath, and survivals of total bacteria and S. aureus were enumerated on tryptic soy agar and mannitol salt agar, respectively, every 30 min for 120 min. At 60oC, the cell counts of total bacteria and the significant difference in survivals between sodium-habituated and non-habituated S. aureus were observed only in the Carbonara sauce; the tailing effect, which is the period of no reduction of bacterial cell counts, was observed in pork cutlet, meat and Carbonara sauces subjected to $60^{\circ}C$. At $70^{\circ}C$, total bacterial populations and sodium-habituated and non-habituated S. aureus cell counts in meat and Carbonara sauce also significantly decreased (p<0.05) after 30 min of heat treatment, followed by the obvious tailing effect. Sodium-habituated S. aureus cell counts in meat and Carbonara sauces were higher (p<0.05) than those of non-habituated S. aureus at $70^{\circ}C$. The results indicate that sodium habituation of S. aureus cells may increase the thermal resistance of the pathogen in RTH sauces; moreover, heating RTH sauces for a short time before serving may not sufficiently decrease the cell counts of S. aureus, particularly for sodium-habituated strain.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.838-843
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• 2006

The chemical processes involved in the formation of green pigment in crushed garlic cloves were investigated based on the principle of pink pigmentation in macerated onions. Intact greening and non-greening garlic cloves were either left untreated or heated at $90^{\circ}C$ for 3 min to inactivate enzyme activities. First, a colorless ether soluble compound referred to as color developer reacted with glycine (among all free amino acids) in garlic to form a second compound insoluble in ether. The latter compound then reacted with formaldehyde to yield the green colored pigment. Alliinase activity was necessary for the production of color developer and for the development of green pigment. In greening garlic that had been heat treated, green pigmentation did not proceed due to the heat-inactivation of alliinase, but the addition of alliinase solution into the garlic homogenates restored the pigmentation. However, this phenomenon was not observed in non-greening garlic with or without heat treatment. Finally, the mechanism of green pigment formation in crushed garlicis similar to that of pink pigment formation in macerated onions.