Proteomic analysis of vitreous biopsy techniques

Jessica M Skeie; Eric N Brown; Harryl D Martinez; Stephen R Russell; Emily S Birkholz; James C Folk; H Culver Boldt; Karen M Gehrs; Edwin M Stone; Michael E Wright; Vinit B Mahajan

doi:10.1097/IAE.0b013e3182562017

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Proteomic analysis of vitreous biopsy techniques

Journal article

Open access

Peer reviewed

Proteomic analysis of vitreous biopsy techniques

Jessica M Skeie, Eric N Brown, Harryl D Martinez, Stephen R Russell, Emily S Birkholz, James C Folk, H Culver Boldt, Karen M Gehrs, Edwin M Stone, Michael E Wright, …

Retina (Philadelphia, Pa.), Vol.32(10), pp.2141-2149

11/2012

DOI: 10.1097/IAE.0b013e3182562017

PMCID: PMC3637028

PMID: 23095728

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https://www.ncbi.nlm.nih.gov/pmc/articles/3637028View

Open Access

Abstract

To compare vitreous biopsy methods using analysis platforms used in proteomics biomarker discovery. Vitreous biopsies from 10 eyes were collected sequentially using a 23-gauge needle and a 23-gauge vitreous cutter instrument. Paired specimens were evaluated by UV absorbance spectroscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and liquid chromatography tandem mass spectrometry (LC-MS/MS). The total protein concentration obtained with a needle and vitrectomy instrument biopsy averaged 1.10 mg/mL (standard error of the mean = 0.35) and 1.13 mg/mL (standard error of the mean = 0.25), respectively. In eight eyes with low or medium viscidity, there was a very high correlation (R = 0.934) between the biopsy methods. When data from 2 eyes with high viscidity vitreous were included, the correlation was reduced (R = 0.704). The molecular weight protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of paired needle and vitreous cutter samples were similar, except for a minority of pairs with single band intensity variance. Using LC-MS/MS, equivalent peptides were identified with similar frequencies (R ≥ 0.90) in paired samples. Proteins and peptides collected from vitreous needle biopsies are nearly equivalent to those obtained from a vitreous cutter instrument. This study suggests both techniques may be used for most proteomic and biomarker discovery studies of vitreoretinal diseases, although a minority of proteins and peptides may differ in concentration.

Biopsy - methods

Biopsy - instrumentation

Electrophoresis, Polyacrylamide Gel

Humans

Middle Aged

Biomarkers - analysis

Male

Eye Proteins - analysis

Spectrophotometry, Ultraviolet

Eye Diseases - diagnosis

Vitreous Body - chemistry

Young Adult

Tandem Mass Spectrometry

Vitrectomy - instrumentation

Adolescent

Proteomics

Chromatography, Liquid

Female

Aged

Details

Title: Subtitle: Proteomic analysis of vitreous biopsy techniques
Creators: Jessica M Skeie - Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA, USA
Eric N Brown
Harryl D Martinez
Stephen R Russell
Emily S Birkholz
James C Folk
H Culver Boldt
Karen M Gehrs
Edwin M Stone
Michael E Wright
Vinit B Mahajan
Resource Type: Journal article
Publication Details: Retina (Philadelphia, Pa.), Vol.32(10), pp.2141-2149
DOI: 10.1097/IAE.0b013e3182562017
PMID: 23095728
PMCID: PMC3637028
NLM abbreviation: Retina
ISSN: 1539-2864
eISSN: 1539-2864
Publisher: United States
Grant note: F32 EY022280 / NEI NIH HHS K08 EY020530 / NEI NIH HHS
Language: English
Date published: 11/2012
Academic Unit: Molecular Physiology and Biophysics; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Ophthalmology and Visual Sciences
Record Identifier: 9983980065402771

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