Methods to determine biotin-binding capacity of streptavidin-coated magnetic particles

Dorgan, Lonnie J.; Magnotti, Ralph A.; Hou, Janming; Engle, Terri; Ruley, Kevin M.; Shull, Bruce

doi:10.1016/s0304-8853(98)00563-0

Cited by 19 publications

(15 citation statements)

References 4 publications

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“…Although the SA±biotin capture technology is widely used in various immunoanalytical systems (Dorgan et al, 1999;Paek et al, 1999;Stenman et al, 1994), problems may arise in a semiquantitative analysis when determined by a bar code signal as dose response. In the preliminary system shown in Figure 1, two dierent antibodies that simultaneously bound to a single analyte molecule were labeled with colloidal gold and covalently coupled to biotin, respectively.…”

Section: Resultsmentioning

confidence: 99%

Semiquantitative, bar code version of immunochromatographic assay system for human serum albumin as model analyte

Cho

Paek

2001

Biotech & Bioengineering

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An immunochromatographic assay system was devised that can express the concentration ranges of analyte (e.g., urinary human serum albumin) as distinct numbers of the ladder bar (bar coding) for semiquantitation. We constructed a model system consisting of five membrane pad strips partially superimposed in a length. Upon wicking of sample from the bottom, the medium dissolved two different biotinylated species, antibody to the analyte and conjugates of the antibody with colloidal gold, and antigen-antibody reactions took place in the hollow space of the glass fiber membrane. After eliminating unreacted biotinylated molecules at the next strip with an immobilized albumin, the immune complexes were transferred to the pad with streptavidin immobilized in a ladder bar pattern. Analytical conditions here were set for competition between the two biotinylated species for the streptavidin binding sites. The degree of such competition was proportional to the analyte concentration and, consequently, the bar signal number was elevated as the concentration increased. Under optimal conditions for sensitivity, the analytical system responded to the analyte doses at between 30 and 120 mg/dL by producing different bar codes within 5 min.

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Section: Resultsmentioning

confidence: 99%

Semiquantitative, bar code version of immunochromatographic assay system for human serum albumin as model analyte

Cho

Paek

2001

Biotech & Bioengineering

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“…Based on these methods, scientists have established enzyme inhibition method [18][19][20] for determining activity of streptavidin and quantified the binding capacity by calculating the amount of biotin that bound with streptavidin. Herein, we used HRP labeled biotin to replace AP labeled biotin as reported in literature to establish a reproducible method.…”

Section: Determination Of Biotin Binding Capacity Of Streptavidin Magmentioning

confidence: 99%

“…Biotin binding capacity of streptavidin GoldMag particles was determined by the enzyme inhibition method [18][19][20] . Briefly, 25 μg of streptavidin GoldMag particles (containing 5 μg of streptavidin) was transferred to 2 mL tubes and treated three times with STE buffer.…”

Section: Biotin Binding Capacity Of Streptavidin Magnetic Particlesmentioning

confidence: 99%

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Preparation and application of streptavidin magnetic particles

et al. 2007

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Two kinds of streptavidin magnetic particles, namely streptavidin GoldMag particles and streptavidin amino terminal particles were prepared by the methods of physical adsorption and covalent interaction respectively. The streptavidin coated on magnetic particle surface, crucial to many applications, was greatly influenced by the choice of the different buffer. Compared with Dynalbeads®M-270 streptavidin, the binding capacity for biotin of different streptavidin magnetic particles was determined by enzyme inhibition method, and the coupling capacity and activity of biotinylated oligonucleotide on their surface were also analyzed. The results indicated that the streptavidin GoldMag particle prepared by physical adsorption was stable in STE (NaCl-Tris-EDTA) buffer that was frequently used in nucleic acid hybridization and detection. The streptavidin amino terminal particles prepared by covalent interaction could be used both in STE buffer and PBS (phosphate buffered saline) buffer. The biotin binding capacity for 1 mg of streptavidin GoldMag particles and streptavidin amino terminal particles was 4950 and 5115 pmol respectively. The capacity of biotinylated oligonucleotide (24 bp) coupled on 1 mg of GoldMag and amino terminal magnetic particles was 2839 and 2978 pmol separately. These data were about 6-7 times higher than those of Dynabeads®M-270 streptavidin. The hybridization results with FITC-labeled complementary probe on magnetic particle surface demonstrated that the oligonucleotide coupled on streptavidin magnetic particles had high biological activity.

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“…Then, this sample is covalently grafted with streptavidin [15]. A number of 185 available binding sites per bead is measured by inverse titration using fluorescein biotin [16]. As one of the most important requirements, this colloidal dispersion is stable and does not aggregate during a storage period of several months or under magnetic field.…”

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confidence: 99%

Magnetic Force Probe for Nanoscale Biomolecules

Koenig

Hébraud²,

Gosse

et al. 2005

Phys. Rev. Lett.

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We present a new technique to measure the mechanical properties of small biomolecules. This technique uses long range repulsive colloidal forces together with magnetic attraction as a force probing tool. The biomolecules are grafted between superparamagnetic particles, which are regularly spaced within long chains maintained by an external magnetic field. Varying the magnetic field results in compression or extension of the molecules between the particles. In order to demonstrate this technique we use, as a size controlled model molecule, a short double stranded DNA (151 base pairs) for which the force-extension law is determined and found in agreement with existing predictions.

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Methods to determine biotin-binding capacity of streptavidin-coated magnetic particles

Cited by 19 publications

References 4 publications

Semiquantitative, bar code version of immunochromatographic assay system for human serum albumin as model analyte

Semiquantitative, bar code version of immunochromatographic assay system for human serum albumin as model analyte

Preparation and application of streptavidin magnetic particles

Magnetic Force Probe for Nanoscale Biomolecules

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