• Archives of Pharmacal Research
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• v.12 no.3
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• pp.186-190
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• 1989

The release rates of methotrexate (MTX) from MTX-human serum albumin (HSA) conjugate, and 5-fluorouracil (5-FU) from 5-FU acetic acid (AA)-HSA conjugate were determined after incubation of the conjugates in various conditions. The concentrations of 5-FU released from the conjugate increased monoexponentially, however those of MTX increased biexponentially in all studies. It indicated that there are two distinct types of MTX-HSA linkage, weakly and tightly bound linkages. The release rates of 5-FU were lower than those of MTX in all studies indicating that the bond of 5-FU-AA-HSA conjugate is very stable, which is supported by the higher value of activation energy (39. 9 vs 10. 7 Kcal/mole) using Arrhenius equation. The release rates of MTX and 5 -FU from the conjugates increased with incubation temperatures. Proteolytic enzyme and liver homogenates accelerated significantly the release rates of MTX and 5-FU. Approximately 1.30 and 22.0% of MTX were released after 12 hours of incubation in the absence and presence of protease, respectively. The corresponding values for 5-FU were released after 12 hours of incubation with rat liver homogenates which were diluted 6 times with phosphate buffer of pH 6.0. The MTX-HSA and 5-FU-AA-HSA conjugates were very stable in rat plasma.

• YAKHAK HOEJI
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• v.33 no.5
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• pp.267-272
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• 1989

The durg-macromolecule conjugates i.e. 5-fluorouracil-1-acetyl-human serum albumin(FU-AA-HSA, 8) and 5-fluorouracil-1-acetyl-poly-L-lysine(FU-AA-polylys, 9) have been synthesized and purified by sephadex G-25 gel filtration with 0.05M phosphate buffer(pH 7.5). The analyses of conjugates gave the molar ratio of FU-AA : HSA of 70-100:1 and FU-AA: poly-L-lysine of 109:1. The weight percent of FU-AA(as $FU-CH_2CO-$) in FU-AA-HSA conjugate was 16-22% and the one in FU-AA-polylys was 22.4%.

• Archives of Pharmacal Research
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• v.16 no.1
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• pp.1-5
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• 1993

The reaction of sulfhydryl groups in human serum ablumin with bacteriostatic and hypotensive notrosating agents such as sodium nitorprusside and sodium nitrite has been examined. The low reactivity of sodium nitroprusside to sulfhydral groups in albumin has been observed and the sterical inaccessilibility of the agent site which sulfhydryl group resides was implicated. The reaction of sodium nitrite with albumin was highly influenced by pH and little reactivity was observed at physiological pH. On the other hand, the reaction between albumin and S-nitrosoglutatione, an intermediate induced from the reaction of glutathione and nitrosating agents, resulted in the rapid decrease of free sulfhydryl groups in albumin. S-Nitrosylation of the sulfhydryl group by S-nitrosoglutathione and the subsequent production of mixed disulfide is the probable route of modification. In the physiological system, S-nitroso-glutathione may act as an active intermediate in expressing reacivity of nitrosating agents to sulfhydryl groups in albumin.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.743-749
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• 2004

The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

• The Korean Journal of Nuclear Medicine
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• v.23 no.1
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• pp.63-69
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• 1989

In order to develop technetium-99m-labelled human albumin microsphere (HAM) for lung scintigraphy, various experimemts such as preparation and fractionation of HAM, establishment of optimal labelling conditions, determination of radiochemical purity, stability test and biodistribution of $^{99m}Tc-HAM$ were carried out. HAM was prepared from the suspension of 1ml aqueous human serum albumin (25%) in 130 ml of olive oil at $130\sim135^{\circ}$ with vigorous stirring. The resulting HAM was fractionated with microsieve to get the desired particle size $(15\sim50\mu)$ and autoclaved for sterilizaiton. The HAM particles were treated with stannous chloride and the pH of the suspension was adjusted to $3.0\sim3.5$ with phosphate buffer. After freeze-drying the contents of single reaction vial containing 5 mg of HAM and 0.2 mg of $SnCl_2$ it was reacted with $Na^{99m}TcO_4$. The labelling yield was higher than 99.5% and the stability of $^{99m}Tc-HAM$ was high enough to maintain 99.1% of radiochemical purity up to 24 hours. Lung and liver uptake in mice was found to be 94% and 0.9%, respectively. Excellent rabbit and human lung scans were also obtained.

• KSBB Journal
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• v.30 no.1
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• pp.44-51
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• 2015

Chinese hamster ovary (CHO) cells are the most widely used mammalian host for the commercial production of recombinant proteins. However, they show relatively low yields of recombinant proteins in comparison with microbial cells. Various strategies have been tried to overcome this drawback. The acetyl moieties are attached to the N-terminus of histone by histone acetyltransferase (HAT) while histone deacetylase (HDAC) removes histone-bound acetyl groups. HDAC inhibitor (HDACi), such as sodium butyrate, sodium propionate and valproic acid, can enhance specific productivity of CHO cells. Human albumin-erythropoietin (Alb-EPO) is a novel 105 kDa protein comprising recombinant human EPO fused to human albumin. In this study, we examined the effects of HDACi on the production of Alb-EPO in CHO cells with various concentrations in the range of 0-1 mM. The results showed that sodium butyrate was found to be the best HDACi for enhancing productivity. It enhanced not only the production of Alb-EPO but also the apoptosis of recombinant CHO cells.

• Clinical and Experimental Reproductive Medicine
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• v.11 no.2
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• pp.69-76
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• 1984

In order to obtain high proportion of Y sperm the semen was laid over 6%,10% and 20% bovine serum albumin. Separated sperm were stained with quinacrine mustard in order to see F - body which could be seen in human Y sperm. But we could'nt find F-body in the bull sperm. So sperm were compared with size of sperm head. As a result of observation separated sperm was small in size of length and width of sperm head as compared with control sperm. So it was found that the proportion of Y sperm showed a marked increase in separated layer. Then the higher albumin density was, the higher the proportion of Y sperm which had smaller head and faster motility was. But the higher albumin density was, the lower the recovery rate of sperm was. So it was hard to separate Y sperm in oligospermia.

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.858-864
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• 2000

The purpose of the present study was to examine the efficacy and mechanism of the fraction IV cold ethanol fractionation and pasteurization ($60^{\circ}C$ heat treatment for 10h) steps, involved in the manufacture of albumin from human plasma, in the removal and/or inactivation of blood-born viruses. A variety of experimental model viruses for human pathogenic viruses, including the Bovine viral diarrhoea virus (BVDV), Bovine herpes virus (BHV), Murine encephalomyocarditis virus (EMCV), and Porcine parvovirus (PPV), 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 then quantified using a 50% tissue culture infectious dose ($TCID_{50}$). The mechanism of reduction for the enveloped viruses (BHV and BVDV) during fraction IV fractionation was inactivation rather than partitioning, however, it was partitioning in the case of the non-enveloped viruses (EMCV and PPV). The log reduction factors achieved during fraction IV fractionation were ${\geq}6.9$ BHV, $\geq5.2$ for BBDV, 4.9 for EMC, and 4.0 for PPV. Pasteurization was found to be a robust and effective step in inactivating the enveloped viruses as well as EMCV. The log reduction factors achieved during pasteurization were $\geq7.0$ for BHV, $\geq6.1$ for BVDV, $\geq6.3$ for EMCV, and 1.7 for PPV. These results indicate that the production process for albumin has sufficient virus-reducing capacity to achieve a high margin for virus safety.

• Archives of Pharmacal Research
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• v.15 no.2
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• pp.162-168
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• 1992

Release characteristics of five different types of hydrophilic albumin microspheres (HAM) containing different ratios of methotrexate-albumin (MTX-HSA) conjugates to free MTX: 1 : 0 (HAMC), 3 :1 (HAMC 3F), 1 :1 (HAMCF), 1:3 (HAMCF3) and 0 : 1 (HAMF) were investigated in the absence or presence of protease using dissolution tester. In all the HAMs studied except HAMC, the MTX was released bi-exponentially in the absence of protease; an initial fast release period up to approximately 6h, and thereafter the release rate was very much slower. The fast release of MTX from the HAMs (such as HAMC3F, HAMCF, HAMCF3 and HAMF) at the initial phase in probably due to the release of "physically associated" MTX from the core of the HAMs. The initial rate constants were 7.2, 8.7, 8.5 and 5.9 times greater than the second rate constants for HAMF, HAMCF3, HAMCF and HAMC3F, respectively. MTX release from HAMC was very slow and mono-phasic. It was at most 2.2% of the total entrapped amount by 24 h. The protease accelerated the release of MTX from the HAMs. The percentages of MTX released from HAMs up to 24 h were 100, 89.0, 75.0, 66.0 and 61.0% for HAMF, HAMCF3, HAMCF, HAMC3F and HAMC, respectively in the presence of protease and the corresponding values in the absence of protease were 30.2 19.0, 10.0, 6.5 and 2.2%, respectively. In vitro release of MTX in the presence of protease varied according to the ratios of MTX-HSA conjugates to MTX; the data set from HAMF, HAMCF3 and HAMCF fits better to monophasic first-order profile more adequately than to zero-order profile, that of HAMC3 monophasic first-order, and that of HAMC to bi-phasic zero-order. Above results suggested that zero-order release rate can be achieved by adjusting the ratio of MTX-HSA conjugates to MTX in the preparation of HAMs such as HAMC3F.as HAMC3F.

• The Korean Journal of Mycology
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• v.6 no.2
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• pp.15-18
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• 1978

The study was performed to investigate the inhibitory action on the growth rate of Candida albicans under the various immunological conditions, Candida albicans was grown in broth media containing 0.1ml. (in total broth volume 4.0ml.) of normal human serum, diabetic patient serum, albumin solution, artifical bulla content and nutrient solution as control subject. The inhibitory action of Candida albicans was indicated by mesurement of transparency rate with use of the spectrophotometer. The results are as follows: 1. Normal human serum shows inhibitory effect on the growth of the candida distinctly. 2. Albumin solution reveals almost similar to that of the effect of normal human serum. 3. Artificial bulla content which obtained by irradiation of ultraviolot ray after application of 1% 8-Methoxy-psolaren cream and the diabetic patient serum shows reduction of inhibitory effect as compared with that of the normal human serum. 4. It is estimated that the titer of negative effect of diabetic patient serum is not related with the variation of immunoglobulin titer in patient serum.