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J Frengen

Publications and source records attributed to J Frengen.

6 recordsLinked to original sources

Fricke gel as a tool for dose distribution verification: optimization and characterization.

With the introduction of conformal techniques in radiation therapy, gel dosimetry plays an important role as a 3D dose verification system. There are two main types of gels in use for dosimetry: Fricke gels and polymer gels. The advantages of polymer gels are improved dose response and stability with no diffusion problems. However, the more complicated fabrication procedure and the greater cost compared to Fricke gels makes polymer gels less attractive in routine clinical use. Dose resolution has recently been introduced as a concept for comparing and optimizing the performance of different types of gel dosimeters. This parameter has not yet been investigated for Fricke gels. In this study, the effect on the dose resolution and the diffusion from different gelatine- and Fe2+-concentrations and different pH was evaluated. Increasing the concentration of gelatine from 6 wt% to 10 wt% influenced the diffusion coefficient the most, while reducing the pH from 2.0 to 1.5 had the largest effect on the dose resolution. For a gel consisting of 10 wt% gelatine, 1.0 mM Fe2+ and pH 1.5 the diffusion coefficient was found to be 1.5 mm2 h-1 and the dose resolution was about 4.1% (at 95% confidence level), for a dose of 40 Gy. By evaluating different dose gradients by the gamma-method, the diffusion was shown to have no clinically relevant impact on the dose distribution and plan acceptance within 3 h of irradiation. The results indicate a potential use of Fricke gels for IMRT verification.

Diffusion↗

Analyte and label binding assay read by flow cytometry.

A new immunometric two-site sandwich assay is introduced, in which a label-scavenging binding partner is added to the sample in addition to the analyte-binding partner. The scavenger binding partner binds excess label antibody, giving a signal proportional to the amount of excess label antibody in the sample solution. A set of two calibration curves is obtained from the two binding partners simultaneously, and a combination of the two signals gives an unambiguous determination of the analyte concentration, even for high analyte concentrations where the hook effect may occur. Two-particle immunofluorometric assays developed for placental alkaline phosphatase and human chorionic gonadotropin on the basis of this principle and yielding signals measured by flow cytometry gave rapid results (2 h) and had working ranges in excess of 5 and 6 orders of magnitude for the respective analytes.

Alkaline Phosphatase↗

A sequential binding assay with a working range extending beyond seven orders of magnitude.

A new immunometric sequential binding assay has been developed in which the sample is first reacted with a solid phase binding partner in low concentration, and subsequently with a second binding partner at a higher concentration. The amounts of analyte bound to the two solid phase binding partners are separately measured, thus establishing a double standard curve. There is a shift between the two standard curves along the concentration axis. Thus an unambiguous determination of analyte concentration is obtained, even in the descending region of the curves where the 'hook' effect causes decreasing signal with increasing analyte concentration. A two-particle immunofluorometric assay for AFP based on this principle measured by flow cytometry, resulted in an assay with rapid binding (approximately 2 h), a detection limit of 0.1 kIU/l and a working range (0.3 to > 3 x 10(6) kIU/l) in excess of 7 log10 orders. Assay results compared well with those of an immunoradiometric assay.

Analysis of Variance↗

Dual analyte assay based on particle types of different size measured by flow cytometry.

Simultaneous flow cytometric assays have been developed for alpha-fetoprotein (AFP) and human chorionic gonadotropin (hCG), with internal determination of sample related non-specific binding (NSB). The assays use particles of 7.5, 6.5 and 5.5 microns diameter coated with, respectively, monoclonal antibodies specific for AFP, hCG or an epitope normally not present in serum. The different particle types were identified simultaneously by light-scatter measurements as their specific immunofluorometric responses were determined. The NSB in the simultaneous assay of AFP and hCG was increased by approximately 30% compared to corresponding single analyte assays. The working range of the dual analyte assays was 0.6-2000 kIU/l for AFP and 6-10,000 IU/l for hCG. No significant interference from the presence of the other analyte was observed in the measurement of either AFP or hCG. The 95% confidence interval for the ratio of dual over single analyte assay results was [0.81, 1.11] for AFP and [0.88, 1.16] for hCG.

Acrylates↗

Demonstration and minimization of serum interference in flow cytometric two-site immunoassays.

The ability of serum factors to cross-link labeled mouse monoclonal antibody (mAb) of irrelevant specificity (mAb FN61, subclass IgG1) to different particle types coated with sheep IgG, bovine gamma-globulin, or mAb FN61 was measured simultaneously by flow cytometry. Significant interference with mAb FN61-coated particles was detected in 53 of 101 sera. Of the 30 sera showing the most pronounced interference, 23 were characterized by an even stronger cross-linking to particles coated with bovine gamma-globulin. These were designated type 1 sera. Seven sera, designated type 2, displayed a dominant interference with the mAb FN61-coated particles. The interference reaction in the two serum types was characterized by different kinetics, dependence on particle concentration, and response to blocking agents. The interference was minimized by addition of 500 micrograms of bovine gamma-globulin and 50 micrograms of mAb HH1 (IgG1) of irrelevant specificity per 10 microL of serum sample in a final assay volume of 100 microL.

Antibodies, Monoclonal↗

Homogeneous immunofluorometric assays of alpha-fetoprotein with macroporous, monosized particles and flow cytometry.

We evaluated two homogeneous immunofluorometric assays (IFMAs) of alpha-fetoprotein (AFP) based on new macroporous acrylate particles combined with flow cytometry. The standard IFMA, requiring 1 h of incubation, provided a working range from 1.8 to > 900 kIU/L (CV < 10%) and a detection limit of 0.6 kIU/L. Use of overnight incubation and a lower particle concentration extended the working range by 1 decade in the lower end. Analytical recoveries for the standard IFMA varied between 97% and 108%. The slope and y-intercept of the regression line correlating measurements by the standard IFMA and a routine immunoradiometric assay were not significantly different from 1 and 0, respectively (P > 0.5), and the correlation coefficient was 0.996. High precision and warning of spuriously high measurements were obtained by including in each sample separate particle types for detecting instrument instability and measuring nonspecific binding only.

Acrylates↗