• Title/Summary/Keyword: affinity matrix

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Cross-linked Leucaena Seed Gum Matrix: An Affinity Chromatography Tool for Galactose-specific Lectins

  • Seshagirirao, Kottapalli;Leelavathi, Chaganti;Sasidhar, Vemula
    • BMB Reports
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    • v.38 no.3
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    • pp.370-372
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    • 2005
  • A cross-linked leucaena (Leucaena leucocephala) seed gum (CLLSG) matrix was prepared for the isolation of galactose-specific lectins by affinity chromatography. The matrix was evaluated for affinity with a known galactose-specific lectin from the seeds of snake gourd (Trichosanthes anguina). The matrix preparation was simple and inexpensive when compared to commercial galactose-specific matrices (i.e. about 1.5 US$/100 ml of matrix). The current method is also useful for the demonstration of the affinity chromatography technique in laboratories. Since leucaena seeds are abundant and inexpensive, and the matrix preparation is easy, CLLSG appears to be a promising tool for the separation of galactose-specific lectins.

Magnifying Block Diagonal Structure for Spectral Clustering (스펙트럼 군집화에서 블록 대각 형태의 유사도 행렬 구성)

  • Heo, Gyeong-Yong;Kim, Kwang-Baek;Woo, Young-Woon
    • Journal of Korea Multimedia Society
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    • v.11 no.9
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    • pp.1302-1309
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    • 2008
  • Traditional clustering methods, like k-means or fuzzy clustering, are prototype-based methods which are applicable only to convex clusters. On the other hand, spectral clustering tries to find clusters only using local similarity information. Its ability to handle concave clusters has gained the popularity recent years together with support vector machine (SVM) which is a kernel-based classification method. However, as is in SVM, the kernel width plays an important role and has a great impact on the result. Several methods are proposed to decide it automatically, it is still determined based on heuristics. In this paper, we proposed an adaptive method deciding the kernel width based on distance histogram. The proposed method is motivated by the fact that the affinity matrix should be formed into a block diagonal matrix to generate the best result. We use the tradition Euclidean distance together with the random walk distance, which make it possible to form a more apparent block diagonal affinity matrix. Experimental results show that the proposed method generates more clear block structured affinity matrix than the existing one does.

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Effect of Avidin and Biotin in Attachment of Human Adipose Stem Cells to Micronized Acellular Dermal Matrix (지방줄기세포가 무세포 분쇄진피기질(Acellular micronized dermal matrix) 부착에 있어 Avidin과 Biotin의 효과)

  • You, Gyeol;Rhie, Jong Won;Lim, Jin Soo
    • Archives of Plastic Surgery
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    • v.35 no.1
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    • pp.1-6
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    • 2008
  • Purpose: In tissue engineering, it is important that the scaffolds have high affinity with cells for making efficient use of cells. The authors studied the binding affinity of human adipose stem cells(ASCs) to micronized acellular dermal matrix(alloderm) using biotin and avidin linkages.Methods: Human ASCs were harvested from adipose tissue obtained by abdominoplasty. ASCs($1{\times}10^4$, $5{\times}10^4$, $1{\times}10^5$, $5{\times}10^5$, $1{\times}10^6$, $5{\times}10^6$ cells) were attached to micronized alloderm(1mg) in three groups; 1) control group in which no ASCs and alloderm was treated; 2) serum group in which alloderm was exposed to fetal bovine serum; and 3) biotin group in which biotinylated cells were attached to biotinylated alloderm. The binding affinities were determined 1 day after making ASC-alloderm complexes. The proliferation rates were determined by XTT assays in 4, 7, 14, and 21 days and scanning electron microscopic examination was performed in 7 and 21 days after culture of ASC-alloderm complexes.Results: The binding affinities of the biotin group were significantly increased in all cell concentrations. Maximum binding affinity was observed at $5{\times}10^4/mg$ of micronized dermal matrix in biotin group. The viabilities were lowest in biotin group in contrast to binding affinity, but the difference was not significant. SEM showed well attachment of cells to micronized dermal matrix in all groups. Conclusion: The use of avidin/biotin facilitated human ASCs attaching to micronized acellular dermal matrix. This attachment would not disturb adipose stem cells viabilities. The present study suggests that avidin/ biotin can be used as making efficient use of cells in adipose tissue engineering.

VARIATIONS OF CONTAMINANT RETARDATION FACTOR IN THE PRESENCE OF TWO MOBILE COLLOIDS

  • Kim, Song-Bae;Kim, Dong-Ju
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2001.09a
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    • pp.115-119
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    • 2001
  • Contaminant retardation factor is derived from the colloidal and contaminant transport equations for a four-phase porous medium: an aqueous phase, two mobile colloidal phases, and a solid matrix. It is assumed that the contaminant sorption to solid matrix and colloidal particles and the colloidal deposition on solid matrix follow the linear isotherms. The behavior of the contaminant retardation factor in response to the change of model parameters is examined employing the experimental data of Magee et al. (1991) and Jenkins and Lion (1993). In the four-phase system, the contaminant retardation factor is determined by both the contaminant association with solid matrix and colloidal particles and the colloidal deposition on solid matrix. The contaminant mobility is enhanced when the affinity of contaminants to mobile colloids increases. In addition, as the affinity of colloids to solid matrix decreases, the contaminant mobility increases.

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Affinity column과 베타수용체 정제조건

  • 고광호;신찬영;강현삼
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.145-145
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    • 1993
  • 일반적으로 단백질의 정제를 위한 가장 효과적인 방법은 affinity chromatography법으로 알려져 있다. 본 연구에서는 베타 수용체의 순수정제를 위한 효과적인 affinity matrix를 제조하고 이의 특징을 확인함으로써 수용체 정제를 위한 기초를 수립하고자 하였다. Bisoxirane reagent인 1,4-butanediol diglycydyl ether를 Sepharose CL-4B gel과 반응시켜 탄노원자 10개 길이의 spacer arm을 부착시켰다. 여기에 sodium persulfate, dithiothreitol을 연속적으로 사용하여 -SH기를 도입시긴 추 free radical 반응을 이용하여 베타수용체 길항제인 alprenolol을 부착시킴으로써 affinity gel을 제조하였다. alprenolol치 부착반응을 위한 최적온도는 4$0^{\circ}C$로 나타났으며, semimicro Kjeldahl 질소 분석법을 적용하여 gel을 분석한 결과 ml당 0.4-0.6 $\mu$ mol의 alprenolol이 결합하였음을 확인하였다.

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Affinity Separations Using Microfabricated Microfluidic Devices: In Situ Photopolymerization and Use in Protein Separations

  • Chen Li;Lee, Wen-Chien;Lee, Kelvin H.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.240-245
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    • 2003
  • The use of microfabricated microfluidic devices offers significant advantages over current technologies including fast analysis time and small reagent requirements. In the context of proteomic research, the possibility of using affinity-based separations for prefractionation of samples using microfluidic devices has significant potential. We demonstrate the use of microscale devices to achieve affinity separations of proteins using a device fabricated from borosilicate glass wafers. Photolithography and wet etching are used to pattern individual glass wafers and the wafers are fusion bonded at 650$^{\circ}C$ to obtain enclosed channels. A polymer has been successfully polymerized in situ and used either as a frit for packing beads or, when derivatized with Cibacron Blue 3GA, as a separation matrix. Both of these technologies are based on in situ UV photopolymerization of glycidyl methacrylate (GMA) and trimethylolpropane trimethacrylate (TRIM) in channels.

The activation of CD99 inhibits cell-extracellular matrix adhesion by suppressing β1 integrin affinity

  • Lee, Kyoung-Jin;Lee, Sun-Hee;Yadav, Birendra Kumar;Ju, Hyun-Mi;Kim, Min-Seo;Park, Jeong-Hyun;Jeoung, Doo-Il;Lee, Han-Soo;Hahn, Jang-Hee
    • BMB Reports
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    • v.45 no.3
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    • pp.159-164
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    • 2012
  • CD99 is known to be involved in the regulation of cell-cell adhesion. However, it remains unclear whether CD99 controls cell-extracellular matrix adhesion. In this study, the effects of CD99 activation on cell-extracellular matrix adhesion were investigated. It was found that engagement of CD99 with the stimulating antibody YG32 downregulated the adhesion of MCF-7 cells to fibronectin, laminin and collagen IV in a dose-dependent manner. The CD99 effect on cell-ECM adhesion was inhibited by overexpression of the dominant negative form of CD99 or CD99 siRNA transfection. Treatment of cells with $Mn^{2+}$ or by ${\beta}_1$ integrin-stimulating antibody restored the inhibitory effect of CD99 on cell-ECM adhesion. Cross-linking CD99 inactivated ${\beta}_1$ integrin through conformational change. CD99 activation caused dephosphorylation at Tyr-397 in FAK, which was restored by the ${\beta}_1$ stimulating antibody. Taken together, these results provide the first evidence that CD99 inhibits cell-extracellular matrix adhesion by suppressing ${\beta}_1$ integrin affinity.

A response matrix method for the refined Analytic Function Expansion Nodal (AFEN) method in the two-dimensional hexagonal geometry and its numerical performance

  • Noh, Jae Man
    • Nuclear Engineering and Technology
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    • v.52 no.11
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    • pp.2422-2430
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    • 2020
  • In order to improve calculational efficiency of the CAPP code in the analysis of the hexagonal reactor core, we have tried to implement a refined AFEN method with transverse gradient basis functions and interface flux moments in the hexagonal geometry. The numerical scheme for the refined AFEN method adopted here is the response matrix method that uses the interface partial currents as nodal unknowns instead of the interface fluxes used in the original AFEN method. Since the response matrix method is single-node based, it has good properties such as good calculational efficiency and parallel computing affinity. Because a refined AFEN method equivalent nonlinear FDM response matrix method tried first could not provide a numerically stable solution, a direct formulation of the refined AFEN response matrix were developed. To show the numerical performance of this response matrix method against the original AFEN method, the numerical error analyses were performed for several benchmark problems including the VVER-440 LWR benchmark problem and the MHTGR-350 HTGR benchmark problem. The results showed a more than three times speedup in computing time for the LWR and HTGR benchmark problems due to good convergence and excellent calculational efficiency of the refined AFEN response matrix method.

Picomolar Scale Determination of Carbohydrates Covalently Immobilized on Activated Beads Using Hydroxyl Functionality

  • Yu, Jae-Hoon;Chun, Sung-Min;Park, Ho-Koon;Park, Yong-Keun;Jeong, Sun-Joo
    • BMB Reports
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    • v.32 no.1
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    • pp.98-102
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    • 1999
  • Since carbohydrates are major mediators in cell-to-cell adhesion and communication, the development of specific and strong binders against them could generate promising therapeutics. As the first step towards that goal, sugar molecules have to be immobilized to be used as an affinity matrix. The amino functionality in sugar is the most active nucleophile for the immobilization, if the amino group is available. An alternative and general method is to use the hydroxyl group as a direct nucleophile, but the quantitation of immobilized hydroxyl groups is not easily done. To overcome this limitation, we have developed a method to immobilize various isomers of monosaccharides with p-nitrophenyl groups to the beads by using their hydroxyl groups. It was found that the amount of immobilized sugar was independent of the structure of the sugar, but was dependent on the number of hydroxyl groups. We also developed a sensitive method to quantify the amount of immobilized sugar at the picomolar scale by utilizing commercially available glycosidases to release a sensitive reporter molecule, p-nitrophenol, and detect it by HPLC. This new technique would allow a facile quantitation method for immobilized sugar molecules, which could be used as the affinity matrix to develop strong binders against biologically important sugars.

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