Covalently Bound Antibody on Polystyrene Latex Beads: Formation, Stability, and Use in Analyses of White Blood Cell Populations

Siiman, Olavi; Burshteyn, Alexander; Insausti, M.

doi:10.1006/jcis.2000.7279

Cited by 24 publications

(9 citation statements)

References 30 publications

(27 reference statements)

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“…There were no significant differences in the amount of fluorescent DNA released in the supernatant between beads exposed to fecal extract or not (Fig. 5A) [42]. Next, to evaluate whether HMGB1 binding ability of DNA beads remain intact under different pH conditions, DNA beads were incubated with HMGB1 at neutral or acidic pH; all three beads (B1, B2 and B3) efficiently captured HMGB1 from solutions in a concentration-dependent manner.…”

Section: Resultsmentioning

confidence: 95%

Sequestering HMGB1 via DNA-Conjugated Beads Ameliorates Murine Colitis

et al. 2014

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Inflammatory bowel disease (IBD) is chronic inflammation of the gastrointestinal tract that affects millions of people worldwide. Although the etiology of IBD is not clear, it is known that products from stressed cells and enteric microbes promote intestinal inflammation. High mobility group box 1 (HMGB1), originally identified as a nuclear DNA binding protein, is a cytokine-like protein mediator implicated in infection, sterile injury, autoimmune disease, and IBD. Elevated levels of HMGB1 have been detected in inflamed human intestinal tissues and in feces of IBD patients and mouse models of colitis. Neutralizing HMGB1 activity by administration of anti-HMGB1 antibodies or HMGB1-specific antagonist improves clinical outcomes in animal models of colitis. Since HMGB1 binds to DNA with high affinity, here we developed a novel strategy to sequester HMGB1 using DNA immobilized on sepharose beads. Screening of DNA-bead constructs revealed that B2 beads, one linear form of DNA conjugated beads, bind HMGB1 with high affinity, capture HMGB1 ex vivo from endotoxin-stimulated RAW 264.7 cell supernatant and from feces of mice with colitis. Oral administration of B2 DNA beads significantly improved body weight, reduced colon injury, and suppressed colonic and circulating cytokine levels in mice with spontaneous colitis (IL-10 knockout) and with dextran sulfate sodium-induced colitis. Thus, DNA beads reduce inflammation by sequestering HMGB1 and may have therapeutic potential for the treatment of IBD.

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Section: Resultsmentioning

confidence: 95%

Sequestering HMGB1 via DNA-Conjugated Beads Ameliorates Murine Colitis

et al. 2014

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“…The immobilization process was mainly deduced from protocols previously published by Nustad et al [12], Quash et al [13], and Siiman et al [14]. For the production of antidigoxigenin (anti-DIG) antibody covered polystyrene beads, carboxylated particles with a diameter of 2.0 μm were used.…”

Section: Methodsmentioning

confidence: 99%

The elastic properties of single double-stranded DNA chains of different lengths as measured with optical tweezers

et al. 2006

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Optical tweezers are microscopic tools with extraordinary precision in the determination of the position (±2 nm) of a colloid (diameter: ∼2.0 μm) in 3D-space and in the measurement of small forces in the range between 0.1 and 100 pN (pN=10−12 N). Experiments are reported in which single double-stranded (ds)-DNA chains of different length [2,000 base pairs (bp), 3,000, 4,000, and 6,000 bp] are spanned between two colloidal particles by use of appropriate molecular linkers. For the forces applied (≤40 pN) a fully reversible and well reproducible force–extension dependence is found. The data can be well described by both the worm-like chain model or by an approach developed by R. G. Winkler. For the resulting persistence length, a pronounced dependence on the ionic concentration in the surrounding medium is found.

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“…[23][24][25] For the preparation carboxylated particles with a diameter of 2.0 mm (Polysciences Europe, Eppelheim, Germany) were used. The beads from a 50 ml bead suspension (26.5 mg/ml) were washed twice with 200 ml of carbonate buffer (100 mM sodium carbonate, 100 mM sodium bicarbonate (pH 9.6)) and three times with 200 ml buffer A (20 mM phosphate buffer (pH 4.5)).…”

Section: Preparation Of Anti-digoxigenin Antibodies Covered Beadsmentioning

confidence: 99%