Immobilization of proteins on magnetic nanoparticles

被引:33
作者
Wang, TH [1 ]
Lee, WC [1 ]
机构
[1] Natl Chung Cheng Univ, Dept Chem Engn, Chiayi 621, Taiwan
关键词
magnetic nanoparticle; protein immobilization; covalent binding; DNA hybridization;
D O I
10.1007/BF02942276
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of N-ethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticle. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. in contrast to EDC, the yields of protein immobilization via the activation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.
引用
收藏
页码:263 / 267
页数:5
相关论文
共 11 条
[1]   Preparation and characterization of YADH-bound magnetic nanoparticles [J].
Chen, DH ;
Liao, MH .
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 2002, 16 (5-6) :283-291
[2]   Preparation and properties of magnetite and polymer magnetite nanoparticles [J].
Dresco, PA ;
Zaitsev, VS ;
Gambino, RJ ;
Chu, B .
LANGMUIR, 1999, 15 (06) :1945-1951
[3]   High-efficiency intracellular magnetic labeling with novel superparamagnetic-tat peptide conjugates [J].
Josephson, L ;
Tung, CH ;
Moore, A ;
Weissleder, R .
BIOCONJUGATE CHEMISTRY, 1999, 10 (02) :186-191
[4]   Immobilization of proteins and enzymes to fine magnetic particles [J].
Koneracká, M ;
Kopcansky, P ;
Antalík, M ;
Timko, M ;
Ramchand, CN ;
Lobo, D ;
Mehta, RV ;
Upadhyay, RV .
JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS, 1999, 201 :427-430
[5]   Immobilization of yeast alcohol dehydrogenase on magnetic nanoparticles for improving its stability [J].
Liao, MH ;
Chen, DH .
BIOTECHNOLOGY LETTERS, 2001, 23 (20) :1723-1727
[6]   ASSESSMENT OF METHODS FOR COVALENT BINDING OF NUCLEIC-ACIDS TO MAGNETIC BEADS, DYNABEADS, AND THE CHARACTERISTICS OF THE BOUND NUCLEIC-ACIDS IN HYBRIDIZATION REACTIONS [J].
LUND, V ;
SCHMID, R ;
RICKWOOD, D ;
HORNES, E .
NUCLEIC ACIDS RESEARCH, 1988, 16 (22) :10861-10880
[7]   Direct binding of protein to magnetic particles [J].
Mehta, RV ;
Upadhyay, RV ;
Charles, SW ;
Ramchand, CN .
BIOTECHNOLOGY TECHNIQUES, 1997, 11 (07) :493-496
[8]   BIOLOGICAL AND BIOMEDICAL ASPECTS OF MAGNETIC FLUID TECHNOLOGY [J].
ROATH, S .
JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS, 1993, 122 (1-3) :329-334
[9]   Surface modification of superparamagnetic nanoparticles (ferrofluid) studied with particle electrophoresis: Application to the specific targeting of cells [J].
Sestier, C ;
Da-Silva, MF ;
Sabolovic, D ;
Roger, J ;
Pons, JN .
ELECTROPHORESIS, 1998, 19 (07) :1220-1226
[10]   ANTIBODY MAGNETITE NANOPARTICLES - IN-VITRO CHARACTERIZATION OF A POTENTIAL TUMOR-SPECIFIC CONTRAST AGENT FOR MAGNETIC-RESONANCE-IMAGING [J].
TIEFENAUER, LX ;
KUHNE, G ;
ANDRES, RY .
BIOCONJUGATE CHEMISTRY, 1993, 4 (05) :347-352