고처리량 단백질 안정성 분석 시스템

1.  Platform (Main system:단백질 열 및 광산란 안정성 분석장비)) &nbsp;&nbsp;1) Instrument &nbsp;&nbsp;&nbsp;&nbsp;- Simultaneous samples per experiment: 96 &nbsp;&nbsp;&nbsp;&nbsp;- Time to read a 96-well sample plate: 1 min &nbsp;&nbsp;&nbsp;&nbsp;- Temperature range: 15–95 °C &nbsp;&nbsp;&nbsp;&nbsp;- Heating rate: 0.1–10 °C/min &nbsp;&nbsp;&nbsp;&nbsp;- Temperature accuracy: ±0.1 °C &nbsp;&nbsp;&nbsp;&nbsp;- Laser classification: Class 1 &nbsp;&nbsp;2) &nbsp;Fluorescence &nbsp;&nbsp;&nbsp;&nbsp;- Parameters determined: Tm, Tonset, Tm of vector payload ejection, rate of isothermal unfolding &nbsp;&nbsp;&nbsp;&nbsp;- Excitation: 280 nm (UV) LED, 470 nm (blue) LED &nbsp;&nbsp;&nbsp;&nbsp;- Detection: Spectrometer at full 250–750 nm spectral range &nbsp;&nbsp;&nbsp;&nbsp;- Compatibility: Protein intrinsic fluorescence, Reporter fluorescence (DSF, nucleotide dye, membrane protein dye) &nbsp;&nbsp;&nbsp;&nbsp;- Sample precision: <2% CV (Tm) &nbsp;&nbsp;&nbsp;&nbsp;- Protein sample concentration: 0.025–300 mg/ml IgG (protein dependent) &nbsp;&nbsp;3) &nbsp;Light scattering &nbsp;&nbsp;&nbsp;&nbsp;- Parameters determined: Tagg, Tsize, rate of isothermal aggregation, Z-average diameter, size distributions, &nbsp;&nbsp;&nbsp;&nbsp;- polydispersity, colloidal stability(kD, B22, G22) &nbsp;&nbsp;&nbsp;&nbsp;- Excitation: 660 nm (red) laser &nbsp;&nbsp;&nbsp;&nbsp;- Detection: Avalanche photodiode &nbsp;&nbsp;&nbsp;&nbsp;- SLS resolution: ≥15 kDa change of mean MW &nbsp;&nbsp;&nbsp;&nbsp;- DLS hydrodynamic diameter range: 0.3–1000 nm &nbsp;&nbsp;&nbsp;&nbsp;- DLS Size accuracy: ±2% &nbsp;&nbsp;&nbsp;&nbsp;- Molecular weight range: 192 Da–25 MDa &nbsp;&nbsp;&nbsp;&nbsp;- Minimum sample concentration: SLS: 0.05–300 mg/mL IgG (protein dependent), DLS: ≥0.1 mg/mL lysozyme  2. Workstation with software (분석용 워크스테이션 및 전용 소프트웨어) &nbsp;&nbsp;1) version 1.2 software &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;- Export graphs to a file or directly to clipboard &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;-&nbsp;Noise filter applied to fluorescence and SLSdata for better data visualization &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;-&nbsp;Sample legends added to data overlay plots

고처리량 형광 및 광산란 분석을 통해 단백질의 열적 안정성(Tm), 응집 온도(Tagg), 입자 크기(Tsize), Z-average diameter, polydispersity, 콜로이드 안정성(kD, B22) 등 핵심 developability 지표를 동일 샘플, 동일 조건에서 동시에 분석 가능함으로서 단백질 안정성 평가에 활용