Development of an indirect competitive ELISA for the determination of papaverine.

Papaverine (1-(3,4-dimethoxybenzyl)-6,7-dimethoxyisoquinoline, PAP) is a member of the benzylisoquinoline sub-group of the opium alkaloids. It has been widely used for treating diseases like pulmonary arterial embolism and renal or biliary colic. In this paper, a specific conjugate of mono-demethylated papaverine-O-carboxylmethyl ether (MDMPAP-O-CME) and bovine serum albumin (BSA) was synthesized and used as the complete antigen (PAP-BSA), with which we successfully obtained a high-titer anti-PAP polyclonal antibody (pAb) by immunization of rabbits.

Biotin-avidin amplified enzyme-linked immunosorbent assay for determination of isoflavone daidzein.

A biotin-avidin amplified enzyme-linked immunosorbent assay (BA-ELISA) method was developed and optimized for the determination of a weakly estrogenic isoflavone daidzein in serum, urine and Puerariae radix. Specific polyclonal antibody was produced against daidzein by immunization of rabbits with a conjugate of 7-O-(carboxymethyl)-daidzein and bovine serum albumin (BSA). The polyclonal antibody showed specific recognition of daidzein, while cross-reactivities to coumarin, 4-hydroxycoumarin, phenol, and other isoflavones such as puerarin and rutin were all lower than 1%.

Direct immunoassay of estriol in pregnancy serum by capillary electrophoresis with laser-induced fluorescence detector.

A simple and sensitive capillary electrophoretic immunoassay (CEIA) was described for the determination of estriol (E(3)) in pregnant women's serum. The method was based on the competitive reaction of fluorescein-labeled E(3) antigen and E(3) with limited amounts of monoclonal antibody. The addition of the thermally reversible hydrogel, poly-N-iso propylacrylamide (pNIPA) in the buffer serving as a replaceable packing material, improved the reproducibility of the method.

A new competitive enzyme-linked immunosorbent assay (ELISA) for determination of estrogenic bisphenols.

Bisphenol A and other hydroxylated diphenylalkanes (generally known as bisphenols) have been identified as potential estrogenic substances. In this paper, a specific polyclonal antibody was produced against these compounds by immunization of rabbits with a conjugate of 4,4-bis (4-hydroxyphenyl) valeric acid and bovine serum albumin (BHPVA-BSA). The polyclonal antibody showed specific recognition of the bisphenol structure, while the cross reactions of other common phenolic compounds such as phenol, hydroquinol and p-hydroxybenzoic acid were all lower than 1%.

Production and characterization of monoclonal antibody against recombinant human erythropoietin.

Objective: To produce specific monoclonal antibody (mAb) against recombinant human erythropoietin (rHuEPO) for development of highly efficient methods for erythropoietin detection in biological fluids.

Methods: rHuEPO was covalently coupled with bovine serum albumin (BSA) and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western blot.

Development and application of a multi-target immunoaffinity column for the selective extraction of natural estrogens from pregnant women's urine samples by capillary electrophoresis.

In this paper, a methodology for the determination of three naturally occurring estrogens (estradiol, estrone and estriol) in pregnant women's urine has been described. The procedure included immunoaffinity column (IAC) extraction of 4 mL of urine sample and subsequent analysis of the extraction by micellar electrokinetic chromatography (MEKC). A multi-target polyclonal antibody that has high affinity to three estrogens was produced.

Study on the interactions between anti-HIV-1 active compounds with trans-activation response RNA by affinity capillary electrophoresis.

The study on the interactions between two anti-human immunodeficiency virus type 1 (anti-HIV-1) active compounds with trans-activation response (TAR) RNA by affinity capillary electrophoresis (ACE) with UV absorbance detection is presented. The results showed that the novel active molecules could interact with TAR RNA and inhibit the reproduce process of HIV-1. The binding constants were estimated by the change of migration time of the analytes through the change of concentrations of TAR RNA in the buffer solution.

Production and characterization of anti-recombinant human erythropoietin (rhEPO) monoclonal antibody.

Anti-rhEPO McAb is valuable for the determination of recombinant human erythropoietin (rhEPO) levels for diagnosis of renal anemia and for doping control analysis. In this paper, anti-rhEPO hybridoma was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from immunized BALB/c mouse, using an enzyme-linked immunosorbent assay (ELISA) method to screen the positive hybridoma. The purified McAb was characterized by ELISA, SDS-PAGE, and Western-blotting.

Determination of morphine by capillary zone electrophoresis immunoassay combined with laser-induced fluorescence detection.

A competitive immunoassay for detecting morphine in bio-samples was established by capillary zone electrophoresis combined with laser-induced fluorescence detection (CZE-LIF). The antigen of morphine was labeled with isothiocyano-fluorescein (FITC) and then incubated with morphine monoclonal antibody and samples. The linear range was 50-1000 ng/mL, which was suitable for clinical and forensic applications.

Direct immunoassay of estrone by capillary electrophoresis with laser-induced fluorescence detection.

An immunoassay for estrone (E(1)) in women's serum, based on the competitive reaction between fluorescein-labeled complete antigen and E(1) with limited amount of anti-estrone monoclonal antibody is described. A thermally reversible hydrogel, poly-N-isopropylacrylamide (pNIPA), was added to the buffer to improve the reproducibility. With a laser-induced fluorescence (LIF) detector, the capillary electrophoretic immunoassay (CEIA) can be applied to determine E(1) at a concentration lower than 19.

Pharmacokinetics and tissue distribution of iv injection of polyphase liposome-encapsulated cisplatin (KM-1) in rats.

Aim: The pharmacokinetics and biodistribution of cisplatin encapsulated in polyphase liposome (KM-1) were compared with those of free drug in rats.

Methods: The platinum levels in serum and normal organs, after a single dose of iv injection of free or encapsulated cisplatin to rats, were determined by induced coupled plasma atomic emission spectrometry.

Results: Serum platinum concentration-time curve after a single iv dose of KM-1 4.

Flow injection determination of gaseous sulfur dioxide with gas permeation denuder-based online sampling and preconcentration.

A simple and novel flow injection method for the determination of gaseous SO(2) is described based on gas permeation denuder (GPD) online sampling and preconcentration. The GPD is easily prepared with poly(vinylidene) difluoride microporous membrane as gas permeable material and two Perspex blocks with smooth and flat interface and rectangular engraved channels of mirror image. The sample gas is on one side of the membrane and phosphate buffer of pH 7.

Cyclic voltammetry characterization of metal complex imprinted polymer.

Polymer capable of specific binding to Cu(2+)-2, 2'-dipyridyl complex was prepared by molecular imprinting technology. The binding specificity of the polymer to the template (Cu(2+)-2, 2'-dipyridyl complex) was investigated by cyclic voltammetric scanning using the carbon paste electrode modified by polymer particles in phosphate buffer solution. Factors that influence rebinding of the imprinted polymer were explored.

Production and characterization of anti-estrone monoclonal antibody.

Objective: Determination of estrone (E1) levels has a significant meaning in evaluating physiological effect and diagnosing some diseases. In order to detect free E1 in biological fluids, a monoclonal antibody specific for E1 was prepared after the complete antigen of E1 was synthesized. The purified monoclonal antibody was fully characterized for later immunoassay.