• Molecular & Cellular Toxicology
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• 제1권2호
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• pp.87-91
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• 2005

Although 2D-PAGE has been widely used as the primary method for protein separation, difficulties in displaying proteins with an extreme values of isoelectric paint (pI), molecular size and hydrophobicity limit the technique. In addition, time consuming steps involving protein transfer and extraction from the gel-pieces can result in sample loss. Here, we describe a novel protein separation technique with capillary size-exclusion chromatography (CSEC) for rapid protein identification from human plasma. The method includes protein fractionation along with molecular size followed by in-solution tryptic digestion and peptide analysis through reversed phase liquid chromatography (RPLC) coupled to nanoflow electrospray-tandem mass spectrometry (ESI-MS/MS). Tryptic peptides are applied an a $100\;{\mu}m\;i.d.{\times}10mm$ length pre-column and then separated on a $75\;{\mu}m{\times}200mm$ analytical column at -100 nL/min flaw rate. Proteins were identified over the wide ranges of pI (3.7-12.3) when this technique was applied to the analysis of $1-2\;{\mu}L$ of human plasma. This gel-free system provides fast fractionation and may be considered a complementary technique to SDS-PAGE in proteomics.

• KSBB Journal
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• 제27권3호
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• pp.177-185
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• 2012

Inonotus obliquus mushroom, which is a fungus belonging to Hymenochaetaceae family, is known to grow on birth trees in colder northern climates and to be a fungal parasite that draws nutrients out of living trees rather than from the ground. For the separation of protein-bound polysaccharide (PBP) from the culture broth and mycelium of Inonotus obliquus, three well known extraction methods namely hot water, ultrasound and microwave were used. The best extraction conditions to separate the PBP (64.94 mg/g) from mycelium by microwave were found to be for 1 hour and $150^{\circ}C$. The possibility for concentration of extracted PBP solution by using membrane was also studied. The extracted PBP solution was concentrated effectively by using an ultrafiltration membrane and the molecular weight cut off (MWCO) is 30 KDa. It was observed that a concentration by the ultrafiltration membrane is essential not only for the development of clean separation technology but also for enhanced production of PBP. As a result, we have shown that PBP in the final concentrated solution showed approximately 10 times higher than that in the crude solution by application of the developed separation systems. The separation yield of PBP was about 89.79% by gel filtration of purification steps and the purified product was confirmed to be PBP by using FT-IR.

• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• Macromolecular Research
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• 제11권1호
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• pp.53-61
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• 2003

A molecular-thermodynamic model is developed for the salt-induced protein precipitation. The protein molecules interact through four intermolecular potentials. An equation of state is derived based on the statistical mechanical perturbation theory with the modified Chiew's equation for the fluid phase, Young's equation for the solid phase as the reference system and a perturbation based on the protein-protein effective two body potential. The equation of state provides an expression for the chemical potential of the protein. In a single protein system, the phase separation is represented by fluid-fluid equilibria. The precipitation behaviors are simulated with the partition coefficient at various salt concentrations and degree of pre-aggregation effect for the protein particles. In a binary protein system, we regard the system as a fluid-solid phase equilibrium. At equilibrium, we compute the reduced osmotic pressure-composition diagram in the diverse protein size difference and salt concentrations.

• 한국막학회:학술대회논문집
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• 한국막학회 2002년도 춘계 총회 및 학술발표회
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• pp.123-125
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• 2002

• 한국응용과학기술학회지
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• 제30권3호
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• pp.551-559
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• 2013

To investigate the adhesion effect of sodium bisulfite content in making blocked isocyanate, wood adhesive PB-1, PB-2, PB-3 and PB-4 containing sodium bisulfite content of 15%, 22.5%, 30% and 37.5% were synthesized respectively. As a result, when the amount of sodium bisulfite increased in adhesives, the tensile strength was found to be proportionally increased. The final adhesive mixtures were manufactured using a two-components system which are prepared by mixing two separate protein and BI solutions due to the precipitate in the adhesives. As PVA was added to adhesives to increase tenacity, the plywood dehiscence phenomenon after cold pressing process was declined. By addition of PVA, the tensile strength was improved up to $6.5{\sim}7kgf/cm^2$ with BI/protein ratio from 1:6 up to 1:8. Phase separation between milk fat and aqueous layer was disappeared after addition of emulsifier, Tween 20. Additon of Tween 20 showed tensile strength up to $5{\sim}6.5 kgf/cm^2$ at NCO/protein ratio of 1:12 ~ 1:14 without phase separation.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.554-557
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• 2005

한외여과분리를 이용해 albumin과 lysozyme의 혼합용액 중 두 단백질을 분리하는데 대한 전기장의 효과를 조사하였다. albumin 용액을 pH7에서 분리한 결과, 전기장의 의해 투과유속은 증가하나 albumin의 여과계수비는 감소하였다. lysozyme 용액을 pH7에서 분리한 결과, 전기장의 의해 투과유속이 소폭 감소하며 여과계수비는 증가함을 볼 수 있었다. 한외여과에서의 lysozyme의 albumin에 대한 선택도는 전기장에 의해 대폭 향상되었다. 전기한외여과법을 사용하여 lysozyme과 albumin을 효율적으로 분리하기 위한 기초를 확립하였으며 기존 한외여과법의 단점을 크게 개선하였다.

• 한국수산과학회지
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• 제37권6호
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• pp.498-504
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• 2004

Removal of waste generated by Paralichthys olivaceus in the seawater aquarium using a foam separator was investigated. Protein concentration without a foam separator continuously increased until 3 days after stocking and reached at 25 mg/L after 5 days stocking, but protein concentration became lower than the initial protein concentration (2.5 mg/L) with a foam separator. The trends of other fish wastes such as ammonia, total suspended solids (TSS) and chemical oxygen demand (COD) were similar to protein. Dissolved oxygen (DO) in the aquarium decreased below 6.0 mg/L without a foam separator, but with a foam separator the average DO in the aquarium was 7.3 mg/L. Foam separation with the increase of superficial air velocity (SAV) was more effective than that with the fixed SAV. This study showed that wastewater. treatment of seawater aquarium using a foam separator is effective method for a fish waste removal and oxygen supply.

• 멤브레인
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• 제15권2호
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• pp.121-131
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• 2005

한외여과를 통하여 난백으로부터 라이소자임을 분리, 정제하는 연구를 수행하였다. 교반식 한외여과장치에 분획 분자량(MWCO)이 각각 10 kDa, 30 kDa 및 100 kDa의 셀룰로즈 재질 분리막을 사용하여 알부민과 라이소자임의 투과실험 데이터를 수집하였다. 단백질 용액은 20 mM, pH 6, 7, 8의 인산염 완충용액에 난단백 함량을 $1\%,\;2\%,\;3\%$ 및 $10\%$ 첨가하여 제조하였으며, 막간압력차(TMP)는 $0.5\~3$ bar의 구간에서 실험을 하였다. 분리막의 분획분자량이 증가하면서 투과유속은 증가하였으며, 분리막에서의 막간압력차가 증가하면서 그리고 공급원액의 난백농도가 감소하면서 투과유속은 증가하였다. 또한, 분리막의 분획분자량이 감소하면서 알부민의 라이소자임에 대한 선택도는 증가하였으며, 모든 분리막에서 pH 와 공급원액의 난백농도 그리고 막간압력차가 증가하면서 라이소자임의 알부민에 대한 선택도는 증가하였다.

• 대한의용생체공학회:의공학회지
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• 제28권5호
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• pp.601-608
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• 2007

2 Dimensional Gel Electrophoresis(2DGE) is an essentialmethodology for analysis on the expression of various proteins. For example, information for the location, mass, expression, size and shape of the proteins obtained by 2DGE can be used for diagnosis, prognosis and biological progress by comparison of patients with the normal persons. Protein spot matching for this purpose is comparative analysis of protein expression pattern for the 2DGE images generated under different conditions. However, visual analysis of protein spots which are more than several hundreds included in a 2DGE image requires long time and heavy effort. Furthermore, geometrical distortion makes the spot matching for the same protein harder. In this paper, an iterative algorithm is introduced for more efficient spot matching. Proposed method is first performing global matching step, which reduces the geometrical difference between the landmarks and the spot to be matched. Thus, movement for a spot is defined by a weighted sum of the movement of the landmark spots. Weight for the summation is defined by the inverse of the distance from the spots to the landmarks. This movement is iteratively performed until the total sum of the difference between the corresponding landmarks is larger than a pre-selected value. Due to local distortion generally occurred in 2DGE images, there are many regions in whichmany spot pairs are miss-matched. In the second stage, the same spot matching algorithm is applied to such local regions with the additional landmarks for those regions. In other words, the same method is applied with the expanded landmark set to which additional landmarks are added. Our proposed algorithm for spot matching empirically proved reliable analysis of protein separation image by producing higher accuracy.