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Biomedical subjects

M Strandh

Publications and source records attributed to M Strandh.

11 recordsLinked to original sources

Analysis of carbohydrates using liquid chromatography--surface plasmon resonance immunosensing systems.

An immunosensing system based on surface plasmon resonance (SPR) was used for on-line detection and characterization of carbohydrate molecules separated by high-performance liquid chromatography. These analytes, with or without serum, were continuously separated and analyzed in the combined liquid chromatography-surface plasmon resonance (LC-SPR) system. By using weak and readily reversible monoclonal antibodies, the SPR system allowed specific on-line monitoring of the substances. To increase the specificity of the immunosensor, nonrelevant antibodies were used as reference in a serial flow cell. The sensitivity of the LC-SPR system was dependent on molecular weight of the carbohydrate, affinity of binding, and design of the sensor.

Carbohydrates↗

Continuous weak-affinity immunosensing.

A multitude of weak biological interactions, either working alone or in concert, occur frequently throughout biological systems. We have used this natural feature of readily reversible interactions as the basis for continuous immunosensing. In a model system, a set of weak monoclonal antibodies directed towards a carbohydrate epitope was studied with the aid of surface plasmon resonance. Because the system requires no regeneration, it can be used as a truly on-line immunosensing device. This principle should have wide application in all areas where there is a need for the continuous evaluation of a molecule.

Antibodies, Monoclonal↗

New approach to steroid separation based on a low affinity IgM antibody.

IgM antibodies are often of low affinity (dissociation constant (Kd) > 10(-5) M) and therefore they are usually neglected as tools in, e.g., immunoassays. Previous studies have shown that low affinity biological interactions can be studied and exploited in affinity chromatography, biosensor technology and capillary electrophoresis. In this study we have demonstrated that IgM can be a useful ligand for analytical separation of antigens in weak affinity chromatography (WAC). A low affinity human monoclonal IgM antibody, directed at digoxin, was produced in a hybridoma cell culture, purified to homogeneity and immobilized onto an HPLC support. The IgM HPLC column displayed specific weak affinity retention in the 0.01-0.1 mM range as evaluated with digoxin and ouabain. The specificity was not affected when samples of ouabain in a crude environment of diluted serum were separated on the IgM column. These findings suggest an approach in immunoadsorbent technology where biomolecules can be analyzed and separated with weak affinity chromatography using IgM as a general affinity ligand.

Antibodies, Monoclonal↗

Studies of interactions with weak affinities and low-molecular-weight compounds using surface plasmon resonance technology.

Interactions between the immobilized weak-affinity monoclonal IgG antibody 39.5, which is specific for the glucose-alpha 1,4-glucose motif, and various oligosaccharides were studied with surface plasmon resonance technology. The antibody was immobilized at high levels on the surface of the sensor chip and different concentrations of the analytes were injected at 25 and 40 degrees C. The 39.5 antibody exhibited specific binding to maltose, tetraglucose and maltotriose, with dissociation constants Kd in the range from 0.07 mM (25 degrees C) to 1.0 mM (40 degrees C). Association and dissociation rate constants (ka and kd) were rapid and baseline was obtained almost immediately after the end of each antigen injection. This excluded the need for a regeneration step but also made calculation of the kinetic values impossible. Owing to the weak affinity and the small size of the analytes (< 1000 Da), a careful design of control surfaces is demanded to exclude artefactual results.

Animals↗

In vivo absorbed dose measurements with mini-TLDs--parameters affecting the reliability.

Mini-TLDs have been proposed and widely used for in vivo measurements of absorbed doses in radionuclide therapies. The present investigation reports in detail on the signal dependence on different parameters and the accuracy of this method. Rodshaped Teflon-imbedded CaSO4:Dy or LiF thermoluminescent dosimeters (TLDs) with dimensions 0.2 x 0.4 x 5 mm3 were prepared from TLD-discs. To remove paraffin from the mini-TLDs after cutting in a microtome the TLDs were Xylene-treated, which does not affect the sensitivity. Irradiated mini-TLDs are sensitive to illumination. Fading effects in darkness were examined after 60Co-irradiation at temperatures 4, 22 and 37 degrees C. For CaSO4:Dy mini-TLDs fading in air is small. The observed signal loss after implanting CaSO4:Dy mini-TLDs in gel and muscle tissue is the same at constant temperature and is increasing with the temperature. For LiF mini-TLDs the effect of signal loss in gel was smaller than for CaSO4:Dy dosimeters. For 60Co external irradiation supralinearity already starts between 0.5 and 1 Gy for both kinds of dosimeter material. There is a strong pH dependence of the signals from the mini-TLDs. For CaSO4:Dy dosimeters the loss of sensitivity in gel is smaller at higher pHs. For LiF dosimeters the loss of sensitivity is smallest for neutral pH. We conclude that using mini-TLDs for in vivo dosimetry requires careful handling and proper calibration for accuracy in the measurements. Without such calibration errors exceeding 65% for CaSO4:Dy and 40% for LiF may easily occur.

Air↗

Parameters to consider for measurements of absorbed doses in vivo with mini-thermoluminescent dosimeters.

BACKGROUND: For systemic radiation therapy, i.e., radioimmunotherapy, there is a demand for direct methods of measuring the absorbed dose in vivo. One such method is the use of mini-thermoluminescent dosimeters (TLDs). This paper reports an investigation of the sensitivity of tissue implanted mini-TLDs (calcium sulfate:Dy, 0.2 x 0.4 x 5.0 mm). METHODS: After being irradiated with cobalt-60, the mini-TLDs were left for as long as 9 days in air, gel, and muscle tissue. RESULTS: There was an extensive signal loss, which increased with time, except in air. After 9 days in gel or muscle tissue at room temperature, the signal was decreased to one third of its original value. The dosimeters needed to be kept in constant darkness. There was a strong pH dependence, with a loss of sensitivity of 63% at a pH below 5, which got smaller at higher pH values and reached 10% at pH = 10. CONCLUSIONS: When using mini-TLDs in vivo, one must calibrate the dosimeters in similar milieus, unless the position of the dosimeters in tissue after implantation can be monitored for temperature, pH, and liquid flow.

Animals↗

Electron microscopy and computed microtomography studies of in vivo implanted mini-TL dosimeters.

The need for direct methods of measuring the absorbed dose in vivo increases for systemic radiation therapy, and in more sophisticated methodologies developed for radioimmunotherapy. One method suggested is the use of mini-thermoluminescent dosimeters (TLD). Recent reports indicate a marked loss of signal when the dosimeters are used in vivo. We investigated the exterior surface of the dosimeters with scanning electron microscopy and the interior dosimeter volume with computed microtomography. The results show that the dosimeters initially have crystals uniformly embedded in the teflon matrix, with some of them directly exposed to the environment. After incubation in gel, holes appear in the dosimeter matrix where the crystals should have been. The computed microtomographic images show that crystals remain in the interior of the matrix, producing the remaining signal. We conclude that these dosimeters should be very carefully handled, and for practical use of mini-TLDs in vivo the dosimeters should be calibrated in equivalent milieus. An alternative solution to the problem of decreased TL efficiency, would be to coat the dosimeters with a thin layer, of Teflon, or other suitable material.

Animals↗

Detection and characterization of weak affinity antibody antigen recognition with biomolecular interaction analysis.

In biological systems, weak-affinity interactions (association constant, Ka, of less than approximately 10(4) M-1) between biomolecules are common and essential to the integrity of such units. However, studies of weak biological interactions are difficult due to the scarcity of analytical methods available for the bioscientist. In this communication, we report on the use of biosensors based on surface plasmon resonance to detect and characterize weak affinity antibody-antigen interactions. Monoclonal antibodies towards carbohydrate antigens were immobilized on sensor surfaces and were used to detect weak binding of the carbohydrate tetraglucose of dissociation constant, Kd, in the millimolar range. Sensorgrams were received in the form of square pulses where the kinetic rate constants were difficult to assess due to the rapid association and dissociation of the antigen to/from the immobilized antibody.

Antibodies, Monoclonal↗

Considerations on the calibration of small thermoluminescent dosimeters used for measurement of beta particle absorbed doses in liquid environments.

An investigation has been carried out on the factors which affect the absolute calibration of thermoluminescent dosimeters (TLDs) used in beta particle absorbed dose evaluations. Four effects on light output (LO) were considered: decay of detector sensitivity with time, finite TLD volume, dose linearity, and energy dependence. Most important of these was the decay of LO with time in culture medium, muscle tissue, and gels. This permanent loss of sensitivity was as large as an order of magnitude over a 21-day interval for the nominally 20-microns-thick disc-shaped CaSO4(Dy) TLDs in gel. Associated leaching of the dosimeter crystals out of the Teflon matrix was observed using scanning electron microscopy. Large channels leading from the outside environment into the TLDs were identified using SEM images. A possibility of batch dependence of fading was indicated. The second most important effect was the apparent reduction of light output due to finite size and increased specific gravity of the dosimeter (volume effect). We estimated this term by calculations as 10% in standard "mini" rods for beta particles from 90Y, but nearly a factor of 3 for 131I beta particles in the same geometry. No significant nonlinearity of the log (light output) with log (absorbed dose) over the range 0.05-20.00 Gy was discovered. Energy dependence of the LO was found to be not detectable, within measurement errors, over the range of 0.60-6.0 MeV mean energy electrons. With careful understanding of these effects, calibration via gel phantom would appear to be an acceptable strategy for mini TLDs used in beta absorbed dose evaluations in media.(ABSTRACT TRUNCATED AT 250 WORDS)

Beta Particles↗