Synthetic Polyclonal-Derived CDR Peptides as an Innovative Strategy in Glaucoma Therapy
Abstract
:1. Introduction
2. Experimental Section
2.1. Retina Isolation and Homogenization
2.2. CDRs and a Scrambled Peptide as Control Peptides
2.3. Identification of CDR-Specific Epitope Targets
2.4. Retinal Explants and Immunohistology
2.5. In-Gel and In-Solution Trypsin Digestion
2.6. LC-MS/MS Analysis
2.7. Protein Identification and Quantification
2.8. Data Analysis and Bioinformatics
3. Results
3.1. Identification of CDR-Specific Epitope Targets
3.2. CDR-Induced Effects in an Ex Vivo Glaucoma Model
3.3. CDR-Induced Proteomic Changes in Retinal Explants
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
Abbreviations
AAB | Autoantibodies |
ABC | Ammonium bicarbonate |
ACN | Acetonitrile |
AGC | Automatic gain control |
AMD | Age-related macular degeneration |
CDR | Complementarity-determining region |
CID | Collision-induced dissociation |
CTRL | Control group |
DE | Dynamic exclusion |
DTT | Dithiothreitol |
EtOH | Ethanol |
FA | Formic Acid |
Fab | Antigen-binding fragment |
Fc | Crystallisable fragment |
FDR | False discovery rate |
FR | Framework region |
IAA | Iodoacetamide |
IOP | Intraocular pressure |
IPA | Ingenuity Pathway Analysis |
LC-MS | Liquid chromatography-mass spectrometry |
MeOH | Methanol |
MS | Mass spectrometry |
ONC | Optic nerve cut |
PBS | Phosphate-buffered saline |
POAG | Primary open-angle glaucoma |
RGC | Retinal ganglion cells |
ROS | Reactive oxygen species |
RPE | Retinal pigment epithelium |
STRING | Search Tool for the Retrieval of Interacting Genes/Proteins |
TIC | Total ion current |
TFA | Trifluoroacetic acid |
VH | Variable heavy chain |
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Canonical Pathway | −Log (p-Value) | Molecules |
---|---|---|
Mitochondrial Dysfunction | 3.37 | COX6C, ATP5MF, VDAC2 |
Protein Ubiquitination Pathway | 2.82 | PSMA6, HSPE1, HSP90AA1 |
Oxidative Phosphorylation | 2.39 | COX6C, ATP5MF |
PI3K/AKT Signaling | 2.24 | YWHAE, HSP90AA1 |
Thioredoxin Pathway | 2.21 | TXN |
Pentose Phosphate Pathway (Non-oxidative) | 2.21 | TKT |
Aldosterone Signaling in Epithelial Cells | 2.03 | HSPE1, HSP90AA1 |
Pentose Phosphate Pathway | 2.02 | TKT |
NRF2-mediated Oxidative Stress Response | 1.89 | ERP29, TXN |
Molecular and Cellular Functions | p-Value | Number of Molecules |
---|---|---|
Post-Translational Modification | 8.59 × 10−7–8.59 × 10−7 | 4 |
Protein Folding | 8.59 × 10−7–8.59 × 10−7 | 4 |
Molecular Transport | 1.31 × 10−2–7.28 × 10−6 | 14 |
Protein Trafficking | 1.25 × 10−2–7.28 × 10−6 | 6 |
Cellular Function and Maintenance | 1.48 × 10−2–3.57 × 10−5 | 9 |
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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Schmelter, C.; Fomo, K.N.; Perumal, N.; Manicam, C.; Bell, K.; Pfeiffer, N.; Grus, F.H. Synthetic Polyclonal-Derived CDR Peptides as an Innovative Strategy in Glaucoma Therapy. J. Clin. Med. 2019, 8, 1222. https://doi.org/10.3390/jcm8081222
Schmelter C, Fomo KN, Perumal N, Manicam C, Bell K, Pfeiffer N, Grus FH. Synthetic Polyclonal-Derived CDR Peptides as an Innovative Strategy in Glaucoma Therapy. Journal of Clinical Medicine. 2019; 8(8):1222. https://doi.org/10.3390/jcm8081222
Chicago/Turabian StyleSchmelter, Carsten, Kristian Nzogang Fomo, Natarajan Perumal, Caroline Manicam, Katharina Bell, Norbert Pfeiffer, and Franz H. Grus. 2019. "Synthetic Polyclonal-Derived CDR Peptides as an Innovative Strategy in Glaucoma Therapy" Journal of Clinical Medicine 8, no. 8: 1222. https://doi.org/10.3390/jcm8081222