Quantification of protein deposits on silicone hydrogel materials using stable-isotopic labeling and multiple reaction monitoring.
This study was designed to use multiple reaction monitoring (MRM) for accurate quantification of contact lens protein deposits. Worn lenses used with a multipurpose disinfecting solution were collected after wear. Individual contact lenses were extracted and then digested with trypsin. MRM in conjunction with stable-isotope-labeled peptide standards was used for protein quantification. The results show that lysozyme was the major protein detected from both lens types. The amount of protein extracted from contact lenses was affected by the lens material. Except for keratin-1 (0.83 ± 0.61 vs 0.77 ± 0.20, p = 0.81) or proline rich protein-4 (0.11 ± 0.04 vs 0.15 ± 0.12, p = 0.97), the amounts of lysozyme, lactoferrin, or lipocalin-1 extracted from balafilcon A lenses (12.9 ± 9.01, 0.84 ± 0.50 or 2.06 ± 1.6, respectively) were significantly higher than that extracted from senofilcon A lenses (0.88 ± 0.13, 0.50 ± 0.10 or 0.27 ± 0.23, respectively) (p < 0.05). The amount of protein extracted from contact lenses was dependent on both the individual wearer and the contact lens material. This may have implications for the development of clinical responses during lens wear for different people and with different types of contact lenses. The use of MRM-MS is a powerful analytical tool for the quantification of specific proteins from single contact lenses after wear.
Brien Holden Vision Institute, Sydney, Australia. n.babaei@brienholdenvision.org
Negar Babaei
NB
Omali
Zhenjun
Z
Zhao
Ling
L
Zhong
Mark J
MJ
Raftery
Hua
H
Zhu
Jerome
J
Ozkan
Mark
M
Willcox
D002853:Chromatography, Liquid / Q000379:methods; D003261:Contact Lenses; D005136:Eye Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D020100:Hydrogels / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods*; D053556:Keratin-1 / Q000302:isolation & purification / Q000378:metabolism; D007781:Lactoferrin / Q000302:isolation & purification / Q000378:metabolism; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods; D009113:Muramidase / Q000302:isolation & purification / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D040901:Proteomics; D020034:Silicone Gels / Q000378:metabolism*; D012828:Silicones / Q000378:metabolism*
D005136:Eye Proteins; D020100:Hydrogels; D053556:Keratin-1; D010455:Peptides; D020034:Silicone Gels; D012828:Silicones; C523195:balafilcon A; C523196:senofilcon A; C042434:tear proteins; D009113:Muramidase; D007781:Lactoferrin