Proteomic sample preparation by microdialysis: easy, speedy, and nonselective.

Microdialysis tools have been developed for parallelized medium exchange designated for sample volumes from 10 to 100 μl, compatible with the microplate format, and guaranteeing maximum recoveries without selectivity. These tools are applicable to both protein and peptide analysis. Moreover, they may be used for binding studies as well as for reconcentration and as unique sample containers for complex operating sequences allowing contemporaneous processing and high throughput.

Long-term serum proteomes are quite similar under high- and low-flux hemodialysis treatment.

Purpose: Aim of the study was to identify long-term differences of middle and high-molecular-weight serum constituents under high- and low-flux hemodialysis treatments. Thus, the entire predialytic serum proteomes had to be analyzed using identical hemodialysis membrane material but with different cut-off values.

Methods And Results: A cross-over study and a global native chromatographic proteomic approach were used to analyze serum compositions of 16 patients suffering from end-stage renal disease.

Spinal antinociceptive effects of cyclooxygenase inhibition during inflammation: Involvement of prostaglandins and endocannabinoids.

Both cyclooxygenase-1 and -2 are expressed in the spinal cord, and the spinal COX product prostaglandin E(2) (PGE(2)) contributes to the generation of central sensitization upon peripheral inflammation. Vice versa spinal COX inhibition is considered an important mechanism of antihyperalgesic pain treatment. Recently, however, COX-2 was shown to be also involved in the metabolism of endocannabinoids.

Searching biomarker candidates in serum using multidimensional native chromatography. I. Enhanced separation method.

The microplate-based method developed by our group for non-denaturing multidimensional proteome separation was improved on using improved column arrays and a newly developed robot. Currently size exclusion, anion exchange and lectin affinity chromatography are combined orthogonally. Different samples run simultaneously to enhance reliability of intercomparison.

Searching biomarker candidates in serum using multidimensional native chromatography. II Method evaluation with Alport syndrome and severe inflammation.

Biomarker search using multidimensional native liquid fractionation of serum in microplates was evaluated. From different donors, homologous sample fractions with UV absorbance depending on state of illness were selected, and their constituents were identified and quantitated by MS. Analysis of sera of patients with Alport syndrome and severe inflammation proved the reliability of the method by confirming characteristic alterations.