Applications to human serums

Davis, B.J. 1964. Disc electrophoresis II Method and application to human serum proteins. Ann. N. Y. Acad. Sci. 121 404-427. 

Purification of serine esterases and proteases application to eel acetylcholinesterase and human serum plasminogen Purification by immuno HPAC application to human serum albumin... 

Nevertheless, manufacturers are interested in the further development of reference materials and reference measurement procedures suited for application in human serum for medical purposes. Therefore, industry appreciates the efforts started by the Joint Committee of Trace-ability and Laboratory Medicine (JCTLM), an initiative supported by the Bureau International des Poids et Mesures (BIPM) and the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) in order to develop suitable reference materials and measurement procedures, as well as to collect information on the activities of reference laboratories. In view of the limited resources and large efforts connected with these activities, clear priorities need to be set. Projects need to take into account the clinical importance of the analyte, consider the technical difficulties that must be overcome, and, most importantly, decide whether improvement of the metrological side is reflected in a gain of medically relevant information. 

A rapid two-dimensional separation of a microspot of protein solution under high field intensity is a new application of the two-dimensional principle. The various fractions emerge as a series of rays out of the application zone and hence the designation star electrophoresis which was introduced by us in connection with work on human serum proteins (P4). A similar discontinuous procedure was already mentioned by Strain in 1951 (S4, S5) for the separation of metal ions and recently used by Pucar (P8) and applied by Keler et al. (K1) to human serum. 

Among others, Wieboldt et al. [57] used a similar technique, immunoaffi-nity ultrafiltration, in conjunction with ion spray HPLC/MS, on a small benzodiazepine library tested for binding to a polyclonal sheep IgG antibody. Dunayevskiy et al. [58] used gel filtration together with CE/MS and LC/MS on a small library of drugs tested for binding to human serum albumin, Chu and coworkers et al. [59-61] reported an application of affinity capillary electrophoresis (ACE)-MS to some tri- and tetrpeptide libraries (>1000 compounds)... 

Only applicable to determine the binding to human serum albumin... 

Application to human urine and human serum. The proposed CE-ECL method was applied to the determination of CIP in urinary samples and blood samples. In this study, both urinary samples and blood samples were spiked with different concentrations levels of CIP. Due to the inherent excellent selectivity and sensitivity of the proposed CE-ECL method, the samples were prepared without extra extraction except some simple procedure such as concentration, filtration, and dilution. Fig. 2(A) was the electropherograms of the blank serum sample (a), and 50 Hg/L CIP standard solution was spiked into serum sample as shown in (b). Fig. 2(B)... 

Once chemoenzymatically produced, these unnatural sialosides have demonstrated utility in a variety of biological applications. A sialoside consisting of 9-0-acetylated sialyl Tn antigen (sTn) was produced by one-pot synthesis and conjugated to human serum albumin to be used to evaluate the role of acetylation in carbohydrate-protein interactions. Para-nitrophenol-tagged sialosides can be used in a colorimetric assay to evaluate sialidase specificity. Biotin-tagged sialosides have been immobilized on a streptavidin-coated surface to study protein-carbohydrate interactions with surface plasmon resonance techniques. Size-defined... 

The applicability of 13C nmr to human serum lipoproteins has been demonstrated (Hamilton et al., 1973, 1974). Spectra and partial peak assignments for high-density (HDL), low-density (LDL), and... 

In closing, we refer to the recent pubHcation [358] in which the synthesis of mesoporous poly-DVB and the characterization of its porous structure were reported. The polymer readily adsorbs another surrogate for P2M. lysozyme of hen egg white with molecular weight of 14 kDa and size of about 4nm, but exhibits a pretty low affinity to human serum albumin. At present, however, it is hard to evaluate whether there is any prospect of real application of the sorbent because no information is available on its hemocompatibihty. 

Applications with sodium cyanoborohydride now include the synthesis of large molecular entities used in biological testing. Conjugation of lactose to human serum albumin by reductive amination (eq 37) and carbohydrate conjugation with PEG amines have been added to the array of transformations reported for NaBHsCN. 

Today, however, GC-GC coupling is seldom used to determine pesticides in environmental samples (2), although comprehensive MDGC has been applied to determine pesticides in more complex samples, such as human serum (19). On the other-hand, new trends in the pesticide market, which is now moving towards the production of optically active enantiomers and away from racemic mixtures, may make this area suitable for GC-GC application. The coupling of non-chiral columns to chiral columns appears to be a suitable solution to the separation problems that such a trend might cause. 

Comprehensive two-dimensional GC has also been employed for the analysis of pesticides from serum, which, although not strictly a forensic analytical problem , provides an example of the promise of this technique to forensic applications, such as the analysis of drugs of abuse (40). Two-dimensional gas chromatograms of a 17-pesticide standard and an extract from human serum are shown in Figure 15.13. The total analysis time of about 5 min, high peak capacity and the separation of all... 

The resolution of these columns for protein mixtures, however, was comparably poor. The peak capacity for human serum albumin was near 3 during 20 min gradient elution. Improvement has been reached by covalent binding of PEI (M = 400-600) onto a 330 A silica of 5 pm particle size [38], The peak capacities of ovalbumin and 2a -arid glycoprotein were 30-40 (tgradienl = 20 min). Enhanced peak capacity and resolution probably were due to the more diffuse structure of PEI coupled to silane moieties than that of strictly adsorbed on silica and cross-linked (see Sect, 2.2). Other applications of covalently adsorbed PEI are discussed in Sect. 4.1. 

There is a wide variety of commercially available chiral stationary phases and mobile phase additives.32 34 Preparative scale separations have been performed on the gram scale.32 Many stationary phases are based on chiral polymers such as cellulose or methacrylate, proteins such as human serum albumin or acid glycoprotein, Pirkle-type phases (often based on amino acids), or cyclodextrins. A typical application of a Pirkle phase column was the use of a N-(3,5-dinitrobenzyl)-a-amino phosphonate to synthesize several functionalized chiral stationary phases to separate enantiomers of... 

Takahashi, N., Takahashi, Y., Ishioka, N., Blumberg, B., and Putnam, F. W., Application of an automated tandem high-performance liquid chromatographic system to peptide mapping of genetic variants of human serum albumin, J. Chromatogr., 359, 181, 1986. 

Urdea, M. S., et al. (1987). A novel method for the rapid detection of specific nucleotide sequences in crude biological samples without blotting or radioactivity application to the analysis of hepatitis B virus in human serum. Gene 61, 253-264. 

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