The Effect of Molecular Weight on Passage of Proteins Through the Blood-Aqueous Barrier

Susanne Ragg, Melissa Key, Fernanda Rankin, Darrell WuDunn

Research output: Contribution to journalArticle

Abstract

Purpose: To determine the effect of molecular weight (MW) on the concentration of plasma-derived proteins in aqueous humor and to estimate the plasma-derived and eye-derived fractions for each protein. Methods: Aqueous humor and plasma samples were obtained during cataract surgery on an institutional review board-approved protocol. Protein concentrations were determined by ELISA and quantitative antibody microarrays. A total of 93 proteins were studied, with most proteins analyzed using 27 to 116 aqueous and 6 to 30 plasma samples. Results: Plasma proteins without evidence of intraocular expression by sequence tags were used to fit a logarithmic model relating aqueous-plasma ratio (AH:PL) to MW. The log(AH:PL) appears to be well predicted by the log(MW) (P < 0.0001), with smaller proteins such as cystatin C (13 kDa) having a higher AH:PL (1:6) than larger proteins such as albumin (66 kDa, 1:300) and complement component 5 (188 kDa, 1:2500). The logarithmic model was used to calculate the eye-derived intraocular fraction (IOF) for each protein. Based on the IOF, 66 proteins could be categorized as plasma-derived (IOF<20), whereas 10 proteins were primarily derived from eye tissue (IOF >80), and 17 proteins had contribution from both plasma and eye tissue (IOF 20-80). Conclusions: Protein concentration of plasma-derived proteins in aqueous is nonlinearly dependent on MW in favor of smaller proteins. Our study demonstrates that for proper interpretation of results, proteomic studies evaluating changes in aqueous humor protein levels should take into account the plasma and eye-derived fractions.

Original languageEnglish (US)
Pages (from-to)1461-1469
Number of pages9
JournalInvestigative ophthalmology & visual science
Volume60
Issue number5
DOIs
StatePublished - Apr 1 2019

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Blood-Aqueous Barrier
Molecular Weight
Aqueous Humor
Proteins
Blood Proteins
Research Ethics Committees
Proteomics
Cataract
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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The Effect of Molecular Weight on Passage of Proteins Through the Blood-Aqueous Barrier. / Ragg, Susanne; Key, Melissa; Rankin, Fernanda; WuDunn, Darrell.

In: Investigative ophthalmology & visual science, Vol. 60, No. 5, 01.04.2019, p. 1461-1469.

Research output: Contribution to journalArticle

Ragg, Susanne ; Key, Melissa ; Rankin, Fernanda ; WuDunn, Darrell. / The Effect of Molecular Weight on Passage of Proteins Through the Blood-Aqueous Barrier. In: Investigative ophthalmology & visual science. 2019 ; Vol. 60, No. 5. pp. 1461-1469.
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