Identification and localization of major soluble vitreous proteins in human ocular tissue

Charlie W. Wu; Jennifer L. Sauter; Patrick K. Johnson; Ci Di Chen; Timothy W. Olsen

doi:10.1016/j.ajo.2003.11.009

Identification and localization of major soluble vitreous proteins in human ocular tissue

Charlie W. Wu, Jennifer L. Sauter, Patrick K. Johnson, Ci Di Chen, Timothy W. Olsen

Ophthalmology

Research output: Contribution to journal › Article › peer-review

41 Scopus citations

Abstract

Purpose To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Design Laboratory investigation. Methods Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectomy. Vitreous specimens were subjected to SDS-PAGE and MALDI-TOF-MS analysis. Six specific antibodies were used to identify proteins using Western blot. Four proteins were localized within ocular tissue in a normal donor eyebank eye. Results We found eight distinct bands on SDS-PAGE in normal vitreous and two additional bands (hemoglobin) in eyes with diabetic retinopathy. Proteins were identified using MALDI-TOF-MS and confirmed by Western blot. We established a quantitative analysis of relative protein concentrations of undiluted vitreous. Immunohistochemistry localized selected proteins within the posterior segment layers. Conclusions We present a normal human vitreous protein profile using current technologies and provide a baseline for comparison to ocular disease states. Tissue distribution of vitreous proteins may help to elucidate more specific protein function.

Original language	English (US)
Pages (from-to)	655-661
Number of pages	7
Journal	American journal of ophthalmology
Volume	137
Issue number	4
DOIs	https://doi.org/10.1016/j.ajo.2003.11.009
State	Published - Apr 2004

ASJC Scopus subject areas

Ophthalmology

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10.1016/j.ajo.2003.11.009

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@article{42f86c31229c40eda038dc5e4be136cd,

title = "Identification and localization of major soluble vitreous proteins in human ocular tissue",

abstract = "Purpose To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Design Laboratory investigation. Methods Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectomy. Vitreous specimens were subjected to SDS-PAGE and MALDI-TOF-MS analysis. Six specific antibodies were used to identify proteins using Western blot. Four proteins were localized within ocular tissue in a normal donor eyebank eye. Results We found eight distinct bands on SDS-PAGE in normal vitreous and two additional bands (hemoglobin) in eyes with diabetic retinopathy. Proteins were identified using MALDI-TOF-MS and confirmed by Western blot. We established a quantitative analysis of relative protein concentrations of undiluted vitreous. Immunohistochemistry localized selected proteins within the posterior segment layers. Conclusions We present a normal human vitreous protein profile using current technologies and provide a baseline for comparison to ocular disease states. Tissue distribution of vitreous proteins may help to elucidate more specific protein function.",

author = "Wu, {Charlie W.} and Sauter, {Jennifer L.} and Johnson, {Patrick K.} and Chen, {Ci Di} and Olsen, {Timothy W.}",

note = "Funding Information: This study was supported by the Minnesota Lions Macular Degeneration Center and the University of Minnesota Vision Foundation, Minneapolis, Minnesota, and by Research to Prevent Blindness, Inc, New York, New York. ",

year = "2004",

month = apr,

doi = "10.1016/j.ajo.2003.11.009",

language = "English (US)",

volume = "137",

pages = "655--661",

journal = "American Journal of Ophthalmology",

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TY - JOUR

T1 - Identification and localization of major soluble vitreous proteins in human ocular tissue

AU - Wu, Charlie W.

AU - Sauter, Jennifer L.

AU - Johnson, Patrick K.

AU - Chen, Ci Di

AU - Olsen, Timothy W.

N1 - Funding Information: This study was supported by the Minnesota Lions Macular Degeneration Center and the University of Minnesota Vision Foundation, Minneapolis, Minnesota, and by Research to Prevent Blindness, Inc, New York, New York.

PY - 2004/4

Y1 - 2004/4

N2 - Purpose To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Design Laboratory investigation. Methods Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectomy. Vitreous specimens were subjected to SDS-PAGE and MALDI-TOF-MS analysis. Six specific antibodies were used to identify proteins using Western blot. Four proteins were localized within ocular tissue in a normal donor eyebank eye. Results We found eight distinct bands on SDS-PAGE in normal vitreous and two additional bands (hemoglobin) in eyes with diabetic retinopathy. Proteins were identified using MALDI-TOF-MS and confirmed by Western blot. We established a quantitative analysis of relative protein concentrations of undiluted vitreous. Immunohistochemistry localized selected proteins within the posterior segment layers. Conclusions We present a normal human vitreous protein profile using current technologies and provide a baseline for comparison to ocular disease states. Tissue distribution of vitreous proteins may help to elucidate more specific protein function.

AB - Purpose To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Design Laboratory investigation. Methods Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectomy. Vitreous specimens were subjected to SDS-PAGE and MALDI-TOF-MS analysis. Six specific antibodies were used to identify proteins using Western blot. Four proteins were localized within ocular tissue in a normal donor eyebank eye. Results We found eight distinct bands on SDS-PAGE in normal vitreous and two additional bands (hemoglobin) in eyes with diabetic retinopathy. Proteins were identified using MALDI-TOF-MS and confirmed by Western blot. We established a quantitative analysis of relative protein concentrations of undiluted vitreous. Immunohistochemistry localized selected proteins within the posterior segment layers. Conclusions We present a normal human vitreous protein profile using current technologies and provide a baseline for comparison to ocular disease states. Tissue distribution of vitreous proteins may help to elucidate more specific protein function.

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Identification and localization of major soluble vitreous proteins in human ocular tissue

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