• Title/Summary/Keyword: human albumin

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Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • Biomedical Science Letters
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    • v.7 no.3
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    • pp.103-110
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    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

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Production of Human Serum Albumin in Chloroplast-Transformed Tobacco Plants

  • Ko, Suk-Min;Kim, Hyun-Chul;Yoo, Byung-Ho;Woo, Je-Wook;Chung, Hwa-Jee;Choi, Dong-Woog;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.233-236
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    • 2006
  • Human serum albumin (HSA) is the most abundant protein in plasma and is the most often used intravenous protein in many human therapies. However, HSA is currently extracted only from plasma because commercially feasible recombinant expression systems are not available. This study attempted to develop an efficient system for recombinant HSA production by chloroplast transformation of tobacco. A HSA cDNA was isolated from a cDNA library constructed with human liver tissue. Chloroplast transformation vectors were constructed by introducing various regulatory elements to HSA regulatory sequences. Vectors were delivered by particle bombardment into leaf explants and chloroplast-transformed plants were subsequently regenerated into whole plants. Southern blot analysis confirmed that the HSA cDNA was incorporated between rps12 and orf70B of the chloroplast genome as designed. Western blot analysis revealed that hyper-expression and increasing the stability of HSA were achieved by modification of the regulatory sequences using the psbA5'UTRs in combination with elements of the 14 N-terminal amino acids of the GFP and the FLAG tag. However, only plants transformed with the vector containing all of these elements were able to accumulate HSA.

Component proteins in cystic fluid of Taenia sodium metacestodes collected surgically from neurocysticercosis patients (인체 유구낭미충증 환자에서 수거한 낭미충 낭액의 성분 단백질의 양상)

  • Yoon Kong;Shin-Yong Kang;Seung-Yull Cho
    • Parasites, Hosts and Diseases
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    • v.28 no.2
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    • pp.101-108
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    • 1990
  • Surgically collected cystic fluid of Taenia solium metacestodes from patients of intracranial cystic lesion were compared in their protein composition with those from naturally infected pigs in Cheju Do, Korea and Ecuador. In non-denaturing discontinuous-polyacryla aide gel electrophoresis (disc-PAGE) , no discernible differences were recognized in banding patterns between the cystic fluids from Cheju Do and Ecuador, and between the cystic quids from pigs and human lesions except wider bands that corresponded to human albumin and T-globulin (in 4 of 9 patients). In reducing SDS-PAGE, bands in the cystic Ruid from Ecuador showed the same banding pattern with that from Cheju Do but two bands of 21 and 17 kDa were stained darker. Cystic quids (rom patients revealed the same protein compositions of the major protein bands of 94, 64, 15, 10 and 7 kDa as in the cystic fluid of pig origin, but human albumin (66 kDa), heavy and light chains of gamma globulin (55 and 22.5 kDa) were contaminated in 4 of 9 cystic fluids. Human CSF proteins seem to have been contaminated during cystic ftuid collection. In any cystic quid from patients, the majcr Protein component was 150 kDa which was subdivided into 15, 10 and 7 kDa in reducing SDS-PAGE.

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Photosensitized oxidative damage of human serum albumin by water-soluble dichlorophosphorus(V) tetraphenylporphyrin

  • Ouyang, Dongyan;Hirakawa, Kazutaka
    • Rapid Communication in Photoscience
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    • v.4 no.2
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    • pp.41-44
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    • 2015
  • Biomolecular photo-damaging activity of a water-soluble cationic porphyrin was examined using human serum albumin (HSA), a water-soluble protein as a target biomolecule model by a fluorometry. Dichlorophosphorus(V) tetraphenylporphyrin ($Cl_2P(V)TPP$), was synthesized and used as a photosensitizer. This porphyrin could bind to HSA and cause the photosensitized oxidation of HSA through the singlet oxygen generation and the oxidative photo-induced electron transfer (ET). Near infrared emission spectroscopy demonstrated the photosensitized singlet oxygen generation by this porphyrin. Decrement of the fluorescence lifetime of $Cl_2P(V)TPP$ by HSA supported the ET mechanism. Furthermore, the estimated Gibb's energy indicated that the ET mechanism is possible in the terms of energy. Because oxygen concentration in cancer cell is relatively low, ET mechanism is considered to be advantageous for photosensitizer of photodynamic therapy.

Characteristics of Protein Chromatography by Affinity Membrane Mudule (친화성 막모듈에 의한 단백질 크로마토그래픽 특성)

  • 이광진;염경호
    • KSBB Journal
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    • v.13 no.2
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    • pp.125-132
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    • 1998
  • Protein affinity membrane was prepared via the coating of chitosan gel on the porous flat polysulfone membrane surface, followed by the immobilization f the reactive dye (Cibacron Blue 3GA) to the chitonsan gel. The maximum protein binding capacity of affinity membrane was about 70${\mu}g/cm^2$ determined by the batch adsorption experiments of human serum albumin (HSA). Using module of this membrane, the characteristics of protein chromatography were investigated through the experiments of elution and frontal chromatography of HSA. This membrane module promises as a chromatography column, since it represented a lower pressure drop and a greater reproducibility. The protein separation ratio was significantly influenced by the flow rate of mobile phase and the injection quantity of HSA. The dynamic protein binding capacity of module decreased from the equilibrium binding capacity with increasing flow rate and approached the value of 15 - 20 ${\mu}g/cm^2$ for flow rates above 6 mL/min.

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An Integrated Process for the Separation and Purification of Biologically Active Proteins from Human Urine (인뇨로부터 유용단백질의 통합 분리정제 공정)

  • 김기용;정광회문흥모
    • KSBB Journal
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    • v.8 no.3
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    • pp.287-294
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    • 1993
  • For the purpose of combining the purification processes for several biologically active proteins form human urine, an efficient integrated fractionation procedure has been investigated. The procedure was started by concentration with ultrafiltration and pH precipitation followed by a selectable combination of chromatography on gel filtration, adsorption, ion exchanger, affinity, and reverse phase column. By this process, the purified urokinase, epidermal growth factor and albumin migrated as a single band on SDS-polyacrylamide gel electrophoresis and were fully active. The recoveries of these purified proteins were 48%, 17%, and 46%, respectively.

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The Effect of Albumin Therapy for Reperfusion Injury Following Transient Focal Cerebral Ischemia in Rats (쥐에서 일과성 국소 뇌허혈 후 생긴 재관류 손상시 알부민치료의 효과)

  • Huh, Pil Woo;Cho, Kyoung Suck;Yoo, Do Sung;Kim, Jae Keon;Kim, Dal Soo;Kang, Joon Ki
    • Journal of Korean Neurosurgical Society
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    • v.30 no.1
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    • pp.12-19
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    • 2001
  • Objective : Albumin is a very useful drug for the improving of cerebral blood volume and the oncotic effect in cerebral ischemia or cerebral vasospasm. The purpose of this study was to examine the morphological and neurological effect of albumin therapy on reperfusion injury following transient focal cerebral ischemia. Materials and Methods : 18 Male Sprague-Dawley rats weighing 270-320g were used. The ischemia model was produced by 2-hour period of transient middle cerebral artery occlusion with a poly-L-lysin coated intraluminal suture. The agent(20% human serum albumin[HSA]) or control solution(NaCl 0.9%) was administered intravenously at a dosage of 1% of body weight immediate after reperfusion following a 2-hour period occlusion. Neurological function was evaluated by the postural reflex and the forlimb placing test during occlusion(at 60 min) and daily for 3 days thereafter. The brain was perfusion-fixed, and infarct volumes and brain edema were measured. Results : The HSA significantly improved the neurological score in treated group. The rats of albumin treatment group showed significantly reduced total infarct volume(by 34%) and brain edema(by 81%) compared with salinetreated rats. Conclusion : HSA showed a substantial effect on the transient focal cerebral ischemia and reperfusion injury model. These results may indicate its usefulness in treating reperfusion injury patients after thrombolysis treatment for the thrombo-embolic major cerebral artery occlusions.

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Enhanced Production of Human Serum Albumin by Fed-Batch Culture of Hansenula polymorpha with High-Purity Oxygen

  • Youn, Jong-Kyu;Shang, Longan;Kim, Moon-Il;Jeong, Chang-Moon;Chang, Ho-Nam;Hahm, Moon-Sun;Rhee, Sang-Ki;Kang, Hyun-Ah
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1534-1538
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    • 2010
  • Fed-batch cultures of Hansenula polymorpha were studied to develop an efficient biosystem to produce recombinant human serum albumin (HSA). To comply with this purpose, we used a high-purity oxygen-supplying strategy to increase the viable cell density in a bioreactor and enhance the production of target protein. A mutant strain, H. polymorpha GOT7, was utilized in this study as a host strain in both 5-l and 30-l scale fermentors. To supply high-purity oxygen into a bioreactor, nearly 100% high-purity oxygen from a commercial bomb or higher than 93% oxygen available in situ from a pressure swing adsorption (PSA) oxygen generator was employed. Under the optimal fermentation of H. polymorpha with highpurity oxygen, the final cell densities and produced HSA concentrations were 24.6 g/l and 5.1 g/l in the 5-l fermentor, and 24.8 g/l and 4.5 g/l in the 30-l fermentor, respectively. These were about 2-10 times higher than those obtained in air-based fed-batch fermentations. The discrepancies between the 5-l and 30-l fermentors with air supply were presumably due to the higher contribution of surface aeration over submerged aeration in the 5-l fermentor. This study, therefore, proved the positive effect of high-purity oxygen in enhancing viable cell density as well as target recombinant protein production in microbial fermentations.