Analysis of human tear proteins by different high-performance liquid chromatographic techniques

Baier, Gottfried; Wollensak, Gregor; Mur, Erich; Redl, Bernhard; Stöffler, Georg; Göttinger, W.

doi:10.1016/s0378-4347(00)83408-8

Cited by 32 publications

(22 citation statements)

References 13 publications

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“…35 Peaks 4,6,8 and 9 showed similar protein patterns and may be different iso-forms of b2-microglobulin.…”

Section: Discussionmentioning

confidence: 95%

“…3 Major tear proteins include lysozyme, lactoferrin, secretory immunoglobulin A (sIgA), serum albumin, lipocalin 4 (previously called tear-specific prealbumin) and lipophilin. 5 The function of the ocular surface in normal and disease states is reflected by tear proteins such as b2-microglobulin (Mw ¼ 11.8 kDa), 6 cystatins (Mw ¼ 14 kDa), 7,8 substance P, 9 epidermal growth factor (EGF), 10 transforming growth factor b1 (TGF-b1) and b2…”

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confidence: 99%

“…Qualitative and quantitative techniques include one-and two-dimensional polyacrylamide gel electrophoresis (PAGE), 8,[21][22][23] isoelectric focusing (IEF), 8,21 crossed immunoelectrophoresis, enzyme-linked immunosorbent assay (ELISA), and high-pressure liquid chromatography (HPLC) techniques such as size-exclusion HPLC, [24][25][26] reversed-phase HPLC, 6 and ion-exchange HPLC. 6 Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry has also been applied to study changes in tear proteins before and after corneal wound healing. 27 More recently, proteomic methods have been used to map tear protein profiles.…”

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confidence: 99%

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Analysis of rabbit tear proteins by high‐pressure liquid chromatography/electrospray ionization mass spectrometry

Zhou

Beuerman

Barathi

et al. 2003

Rapid Comm Mass Spectrometry

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The aim of this study was to develop a fast and reliable analytical procedure for the display of the protein components of tears that can be used to differentiate the status of the ocular surface. Using this new procedure, we analyzed the tear protein components following a corneal wound in the rabbit. Calibrated 10-microL glass, fire-polished capillary micropipettes were used to collect tears from New Zealand White rabbits prior to and daily for 9 days following a unilateral 6-mm diameter centrally placed anterior keratectomy. Tear proteins were eluted by a reversed-phase high-performance liquid chromatography (RP-HPLC) column and the tear protein profile was monitored by electrospray ionization (ESI) mass spectrometry positive total ion current (TIC) chromatography. Tear proteins were reliably separated into 17 peaks, each of which contained one or a number of protein components. The molecular weight of each protein component was determined by on-line ESI. Major tear protein components, lactoferrin, lysozyme (minimally detectable in rabbit tears), albumin, lipocalin, lipophilin and beta2-microglobulin, were tentatively identified by this method. Based on the mass spectrometric data, beta2-microglobulin was found to be glycosylated with N-acetylhexosamine. ESI-positive TIC chromatograms and mass spectra revealed comparative differences in the tear protein spectra after corneal wounding. One day after wounding, rabbit lysozyme with a molecular weight of 14,717 Da was found to be 8-fold higher in the tears of wounded eyes when compared with tears from unwounded eyes. It dropped back to normal 3 days after wounding. The expression of an unidentified tear protein with the molecular weight of 16,060 Da was also elevated after corneal wounding and returned to normal level by day 5. In this study, LC/ESI-MS was developed as a fast, reproducible and simple method for the identification and analysis of many of the protein components of the tears. Importantly, this technique also allows quantification of each component resolved in the chromatogram. This method is very suitable for mapping peptides and proteins (<80 kDa) in tears.

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“…35 Peaks 4,6,8 and 9 showed similar protein patterns and may be different iso-forms of b2-microglobulin.…”

Section: Discussionmentioning

confidence: 95%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Analysis of rabbit tear proteins by high‐pressure liquid chromatography/electrospray ionization mass spectrometry

Zhou

Beuerman

Barathi

et al. 2003

Rapid Comm Mass Spectrometry

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show abstract

“…However, a key difference between transferrin and lactoferrin is that transferrin is acidic and lactoferrin is basic (pI Å 7.8). Lactoferrin is more hydrophobic than transferrin (60). From Fig.…”

Section: Hill Coefficients Collected From Figs 2-5mentioning

confidence: 93%

Cooperative Adsorption of Proteins onto Hydroxyapatite

Luo

Andrade

1998

Journal of Colloid and Interface Science

125

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“…Bisherige Untersuchungen in der Literatur beziehen sich auf den Totalen Antioxidativen Status [10], auf Proteinbestimmungen [1,5,8,13], Enzymbestimmungen [2], auf wasserlösliche Antioxidantien wie z. B. Ascorbinsäure, Tyrosin, Cystein, Glutathion, Harnsäure mittels HPLC [3] bzw.…”

Section: Introductionunclassified

Eine neue Methode zur Bestimmung der antioxidativen Kapazität in der Tränenflüssigkeit

Griebenow

Rieger

Horwath‐Winter³

2003

Spektrum Augeheilkd

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Analysis of human tear proteins by different high-performance liquid chromatographic techniques

Cited by 32 publications

References 13 publications

Analysis of rabbit tear proteins by high‐pressure liquid chromatography/electrospray ionization mass spectrometry

Analysis of rabbit tear proteins by high‐pressure liquid chromatography/electrospray ionization mass spectrometry

Cooperative Adsorption of Proteins onto Hydroxyapatite

Eine neue Methode zur Bestimmung der antioxidativen Kapazität in der Tränenflüssigkeit

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