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Poster Presentations
Day 2, June 23（Mon.）
Room P (Maesato East, Foyer, Ocean Wing)
- 2P-AM-51
Characterization of lentiviral vector proteins by LC-MS/MS
(1Sumitomo Pharma, 2Osaka Univ., 3Thermo Fisher Scientific)
oSatomi Kasahara1,2, Yuki Yamaguchi2, Shio Watanabe3, Daisuke Higo3, Tetsuo Torisu2, Susumu Uchiyama2
Lentiviral vectors (LVVs) are widely used in gene therapy due to their ability to efficiently transduce both dividing and non-dividing cells, enabling stable gene integration. LVVs are enveloped viruses encapsidating two copies of single-stranded RNA and composed of structural and non-structural proteins. During production, LVVs incorporate not only viral proteins but also host cell proteins (HCPs), which may impact vector efficacy and safety. However, the specific effects of HCPs on LVV performance remain largely unexplored.
In this study, LVV samples from two different suppliers were analyzed using SDS-PAGE and mass spectrometry-based proteomics. SDS-PAGE revealed protein bands across a broad molecular weight range, with distinct intensity differences between the samples. Shotgun proteomics identified about 4,000 proteins in Sample 1 and about 6,000 in Sample 2, with most proteins being HCPs. Several cytoplasmic and plasma membrane-associated proteins were commonly detected, suggesting HCP incorporation during vector assembly. However, distinguishing particle-associated HCPs from external contaminants remains challenging. Developing strategies to differentiate these proteins will enhance our understanding of LVV composition and contribute to improving vector purification and manufacturing processes.