A. Bult

Self-assembled Monolayers for Biosensors

The use of self-assembled monolayers (SAMs) in various fields of research is rapidly growing. In particular, many biomedical fields apply SAMs as an interface-layer between a metal surface and a solution or vapour. This review summarises methods for the formation of SAMs upon the most commonly used materials and techniques used for monolayer characterisation. Emphasis will lie on uniform, mixed and functionalised monolayers applied for immobilisation of biological components including (oligo-)nucleotides, proteins, antibodies and receptors as well as polymers. The application of SAMs in todays research, together with some applications will be discussed.

Cyclodextrins in the Pharmaceutical Field

AbstractCyclodextrins (CyDs) are cyclic oligosaccharides, containing a minimum of six D-(+)-glycopyranose units attached by α-1, 4-linkages produced by the action of the cyclodextrin-trans-glycosidase enzyme on a medium containing starch. CyDs are somewhat cone-shaped. The outside of CyDs is hydrophilic and the inside of the cavity is hydrophobic in character. If a molecule fits entirely or at least partially into the cavity, an inclusion complex may be formed. In general, hydrophobic molecules, rather than hydrophilic ones, have a higher affinity to the CyD cavity in aqueous solutions. The CyD complexes thus formed are stabilized by various intermolecular forces, such as hydrophobic interaction, van der Waals forces, hydrogen bonding, release of high energy water molecules in complex formation and release of strain energy in the macromolecular CyD ring. Orally administered CyDs have shown to be harmless, because insignificant amounts are absorbed.Parenterally administered natural CyDs may cause severe ne...

Liposomes and immunoassays

Various aspects of the application of liposomes as a label in immunoassays are reviewed. Methods for the preparation of liposomes, from the basic film method to the more advanced dehydration-rehydration method, are discussed. Furthermore, the markers used in liposome labels, as well as the methods to conjugate liposomes to antigens or antibodies, are summarized. Liposome immunoassays are applied as homogeneous or heterogeneous assays. Homogeneous assays often rely on the lytic activity of complement on antibody-associated liposomes. Another group of homogeneous assays utilizes the inhibitory action of antibodies on the activity of conjugates of mellitin (a bee venom protein) with a hapten. Free mellitin conjugates are able to lyse liposomes effectively. Heterogeneous liposome immunoassays, performed either competitively or non-competitively, resemble more closely standard enzyme linked immunosorbent assays, with the enzyme being replaced by a liposome label. Washing steps are used to separate antigen-specifically bound liposomes from unbound liposomes. All bound liposomes are lysed with a detergent, giving an instantaneous amplification. Flow-injection liposome immunoassays and liposome immunosensors are also described as examples of other possible immunoassay formats.

Derivatization trends in capillary electrophoresis

This survey gives an overview of recent derivatization protocols, starting from 1996, in combination with capillary electrophoresis (CE). Derivatization is mainly used for enhancing the detection sensitivity of CE, especially in combination with laser‐induced fluorescence. Derivatization procedures are classified in tables in pre‐, on‐ and postcapillary arrangements and, more specifically, arranged into functional groups being derivatized. The amine and reducing ends of saccharides are reported most frequently, but examples are also given for derivatization of thiols, hydroxyl, carboxylic, and carbonyl groups, and inorganic ions. Other reasons for derivatization concern indirect chiral separations, enhancing electrospray characteristics, or incorporation of a suitable charge into the analytes. Special attention is paid to the increasing field of research using on‐line precapillary derivatization with CE and microdialysis for in vivo monitoring of neurotransmitter concentrations. The on‐capillary derivatization can be divided in several approaches, such as the at‐inlet, zone‐passing and throughout method. The postcapillary mode is represented by gap designs, and membrane reactors, but especially the combination of separation, derivatization and detection on a chip is a new emerging field of research. This review, which can be seen as a sequel to our earlier reported review covering the years 1991—1995, gives an impression of current derivatization applications and highlights new developments in this field.

Chemiluminescence and immunoassays

The principles of chemiluminescence and the application of chemiluminescent labels and substrates in immunoassays are reviewed. Various immunoassay formats and all known classes of chemiluminescent molecules, including 1,2-dioxetanes, luminol and derivatives, acridinium esters, oxalate esters and firefly luciferins are described as well as the many sensitizers and fluorescent enhancers. Recent promising developments are discussed.

On-line sample preconcentration in capillary electrophoresis, focused on the determination of proteins and peptides.

This overview highlights the possibilities of on‐ or in‐line preconcentration procedures in combination with a CZE separation, focused on the determination of peptides and proteins. The discussed methods, including sample stacking, field‐amplified injection, isotachophoresis, solid phase extraction, membrane preconcentration, electroextraction, supported liquid membranes, hollow fibers, immunoaffinity, and molecularly imprinted polymers technology preconcentration are categorized in electrophoresis‐based and chromatography‐based preconcentration. The chromatography‐based preconcentration is subdivided in low‐specificity and high‐specificity methods. A number of preconcentration methods are available, however, this paper demonstrates that various compounds in different media (aqueous solutions, urine, and plasma) require different preconcentration systems. The preconcentration techniques of first choice in general seem to be solid‐phase extraction and membrane preconcentration, because of their high concentration ability, multiapplicability, relative simplicity and clean‐up capability. For the future, hollow fibers seem to hold a great potential as preconcentration technique, yielding high concentration factors, using simple designs. New techniques, such as hollow fibers, molecularly imprinted polymers technology and supported liquid membranes may have the potential to supersede the conventional preconcentration techniques in some cases. The larger the arsenal of preconcentration techniques becomes, the more efficiently peptides and proteins may be analyzed in the future. These techniques, in some cases, require pre‐cleanup procedures, to ensure the purity of the samples to concentrate.

Penetration of zidovudine into the cerebrospinal fluid of patients infected with HIV

Objective:To investigate penetration of zidovudine (ZDV) into the cerebrospinal fluid (CSF) of HIV-infected patients for whom a lumbar puncture was indicated. Design:A prospective study. Setting:General 525-bed hospital with special funding for treatment and research of HIV-infected patients Patients, participants:Thirty-nine patients with a medical indication for lumbar puncture who used ZDV chronically were included in this study (50 samples in total). Main outcome measure:Determination of ZDV and proteins in CSF and plasma samples. Results:CSF concentrations of ZDV showed little fluctuation 1–8 h after the last ingestion of ZDV. In contrast, plasma levels displayed large variability in this period and decreased exponentially over time. As a result, the CSF/plasma ratio increased linearly over time. No significant relation between the ZDV dose, neither the medical indication for lumbar puncture nor the protein ratio (as a measure for the integrity of the blood–brain barrier), and CSF levels of ZDV was found. The CSF/plasma ratio of ZDV did not give essential information on drug distribution into CSF. Conclusions:Penetration of ZDV into the CSF appears to be independent of the dose (range, 200–1250 mg daily), which may be an explanation for the efficacy of low doses of ZDV in the prevention and treatment of HIV-related neurological diseases. ZDV levels were at steady-state during the first 6 h after ingestion. The CSF/plasma ratio of ZDV concentrations is not an appropriate marker for drug penetration into CSF. AIDS 1993, 7:1581–1587

Pharmacokinetics and metabolism of docetaxel administered as a 1-h intravenous infusion.

Abstract Docetaxel, a taxane antitumor agent, was administered to 24 patients by a 1-h intravenous infusion at a dose level of 100 mg/m2 with pharmacokinetic monitoring. The plasma concentration-versus-time data were fitted with a three-compartment model. The mean area under the curve (AUC) for docetaxel was 3.1 ± 0.9 h · mg/l and the clearance was 34.8 ± 9.3 l/h per m2. There was considerable interpatient pharmacokinetic variability. In 33% of the patient population, metabolites were detected in plasma samples collected 5–30 min after the end of the infusion. The cyclized oxazolidinedione metabolite M4 was most frequently present and was detected in 8 out of 24 patients with maximal concentrations between 0.022 and 0.23 mg/l. Logistic regression analysis was performed to predict M4 docetaxel metabolism. In the final model, alanine aminotransferase and alkaline phosphatase levels were the strongest predictors. No relationship was found between M4 metabolism and percentage decrease in neutrophil count in this study. Three patients with high M4 concentrations in plasma during course 1 suffered from most pronounced fluid retention (grade 2–3) after two to five courses.

Pharmaceutical development of anticancer agents derived from marine sources.

The marine ecosystem is more and more acknowledged as a source of potential anticancer agents. After the identification of a potential substance several hurdles have to be overcome before a marine candidate can enter the clinic. Amongst these are the establishment of a method which ensures sufficient supply and, which is the focus of this review, the development of a clinically useful pharmaceutical formulation. General issues with respect to the pharmaceutical development of marine anticancer agents will be discussed, which will be illustrated by highlighting aspects of the pharmaceutical development and clinical use of some representative compounds.

Anthracycline antitumour agents

A review of physicochemical and analytical properties of anthracycline antitumour agents is presented. The following subjects are discussed: protolytic equilibria, partition and partition coefficients, self-association, adsorptive properties, metal complexation, spectroscopy and chromatography. Furthermore, the stability of anthracyclines in solutions, in pharmaceutical preparations and in biological media is discussed.