Analysis of normal human aqueous humour proteins by two‐dimensional gel electrophoresis

Segers, J.; Rabaey, M; Oye, R. Van

doi:10.1002/elps.1150050109

Cited by 10 publications

(3 citation statements)

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“…AH has been considered as an ultrafiltrate of plasma [56]. The presence of apolipoprotein A1 in AH is, therefore, not unexpected, and has been reported previously [67]. In some instances a slightly larger molecule (MI 28180) was found to comigrate.…”

Section: -3)mentioning

confidence: 83%

“…Due to the similarity in the z/m ratio in the liquid phase and m/z ratio in the gas phase the proteins were thought to belong to the same class of compounds and tentatively identified as fragments of immunoglobulins. The eye is considered an immunologically privileged organ and various immunoglobulins (IgA, IgM, IgG) have been previously detected [56,67,681.…”

Section: -3)mentioning

confidence: 99%

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Comparison of protein mixtures in aqueous humor by membrane preconcentration ‐ capillary electrophoresis ‐ mass spectrometry

et al. 1998

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The significance of proteomic research is coupled with the recent exponential growth of these investigations. Currently, the most popular techniques used for these studies include the coupling of 1- and 2-dimensional electrophoresis with mass spectrometric analysis of the extracted and digested proteins. However, detection limits of gel staining methods have led to a search for complimentary techniques that afford the detection of lower concentrations of biologically relevant proteins. In the present studies, we have evaluated the applicability of on-line capillary electrophoresis - mass spectrometry (CE-MS) for this application. Specifically, we used membrane preconcentration-CE-MS (mPC-CE-MS) to analyze 13 samples of human aqueous humor (AH) from patients with various ocular pathologies (cataract, cataract plus glaucoma, and cataract plus pseudoexfoliation syndrome). This approach enabled rapid analysis of a relatively large volume (1 microL of each specimen, and a protein map for each was created. Measured average molecular weights (Mr) were used to tentatively identify proteins after search of the SWISS-PROT database using TagIdent from ExPaSy. Among those proteins tentatively identified are beta-2 microglobulin (Mr 11731.2), apolipoprotein A1 (Mr 28078.6) and serum albumin (Mr 66400). Proteins with Mr of 4349 (unidentified), 11731.2 (beta-2 microglobulin), 13400-14100 (immunoglobulin fragments), 28078.2 (apolipoprotein A1) and approximately 68000 (serum albumin) were observed in the majority of specimens. Generally no significant differences were noted in the protein composition of aqueous humor samples from different pathologies. However, the absence of an Mr 13345 protein and its oxidized form (Mr 13361) in samples from patients with pseudoexfoliation syndrome was noted. Occasionally the alpha-and beta-chains of hemoglobin, a contaminant in aqueous humor introduced during sampling, were also detected. We conclude from these studies that mPC-CE-MS is an attractive complimentary technique for proteome research, as this approach enables direct mapping and characterization of low concentrations of proteins that are present in complex physiologically derived fluids.

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Section: -3)mentioning

confidence: 83%

Section: -3)mentioning

confidence: 99%

Comparison of protein mixtures in aqueous humor by membrane preconcentration ‐ capillary electrophoresis ‐ mass spectrometry

et al. 1998

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“…Sensitive techniques for protein separation have been used during recent years to analyze the low protein content of intraocular fluids (for review, .see Bito 1985). Such studies have been performed on aqueous humor (Saari et al 1983;Segers et al 1984;Tai-Chin & Okamura 1987) and few have used samples from the vitreous (van Bockxmeer et al 1983;Weller et al 1988). Since the vitrous is bordered by the retina over a large area it is probable that major changes of the retina are reflected within the vitreous (for review see, Balazs & Den-814 linger 1984).…”

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confidence: 99%

Changes in the soluble protein of the human vitreous in vitreoretinal disease

Sjöstrand

Karlsson

Andersson

1992

Acta Ophthalmologica

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Samples of the vitreous were analysed in order to identify changes of soluble proteins in vitreo-retinal disease. The soluble proteins of the vitreous were separated on an anion exchange column (Mono-Q). The degree of neutral proteolytic activity in vitreous body was also measured. The vitreous from cataract cases without vitreoretinal disease was characterized by its low content of soluble proteins equivalent to about 1% of that of serum. Albumin and transferrin were the major identified components and their concentrations were approximately 0.85 and 0.03 g/l, respectively. In cases with vitreoretinal disease the vitreous showed changes of total soluble protein and the appearance of additional protein peaks. In patients with PVR the albumin concentration in the vitreous was found to be three times higher as compared to the control group consisting of patients with cataract. Neutral proteolytic activity in the vitreous was relatively low in both normal and pathological vitreous.

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Two‐dimensional electrophoresis of dog bronchoalveolar lavage fluid proteins

et al. 1990

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Proteins of dog bronchoalveolar lavage fluid, obtained by washing the epithelial lining layer of lungs with phosphate-buffered saline, were separated by two-dimensional electrophoresis. Due to the low protein and high salt content of the bronchoalveolar lavage fluid, samples had to be concentrated and desalted. Following electrophoresis the protein spots were visualized by silver staining. Comparing the two-dimensional protein patterns of bronchoalveolar lavage fluid with that from serum, several lung-specific proteins were detected. The most prominent protein, most probably a surfactant-associated protein, showed isoforms with isoelectric points in the range of pH 4.2-4.8, and a molecular mass of 32 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis after reduction with dithiothreitol.

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Analysis of normal human aqueous humour proteins by two‐dimensional gel electrophoresis

Cited by 10 publications

References 8 publications

Comparison of protein mixtures in aqueous humor by membrane preconcentration ‐ capillary electrophoresis ‐ mass spectrometry

Comparison of protein mixtures in aqueous humor by membrane preconcentration ‐ capillary electrophoresis ‐ mass spectrometry

Changes in the soluble protein of the human vitreous in vitreoretinal disease

Two‐dimensional electrophoresis of dog bronchoalveolar lavage fluid proteins

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