Unchained Labs Aunty High-Throughput Protein Stability Analyzer

Overview

The Unchained Labs Aunty High-Throughput Protein Stability Analyzer is an integrated, benchtop instrument engineered for quantitative, multi-parameter characterization of protein conformational and colloidal stability under thermal stress. It simultaneously applies three orthogonal biophysical techniques—Differential Scanning Full-Spectrum Fluorescence (DSF), Static Light Scattering (SLS), and Dynamic Light Scattering (DLS)—within a single 96-well run. DSF monitors intrinsic tryptophan/tyrosine fluorescence across a continuous 300–650 nm spectral range during controlled thermal ramping (typically 1–5 °C/min), enabling precise determination of melting temperature (Tm) and onset temperature (Tonset). SLS quantifies changes in total scattered intensity at fixed angles to detect early-stage aggregation onset (Tagg), while DLS tracks hydrodynamic radius (Rh) evolution in real time to resolve aggregate growth kinetics (Tsize) and polydispersity shifts. This concurrent acquisition eliminates inter-run variability and provides correlative stability fingerprints essential for formulation development, biosimilar comparability, and upstream process optimization.

Key Features

• True simultaneous triple-mode detection: DSF, SLS, and DLS acquired in parallel—not sequentially—ensuring temporal alignment of conformational, thermodynamic, and colloidal events.
• Ultra-low sample consumption: 8 µL per well enables screening of scarce or expensive candidates (e.g., membrane proteins, viral vectors, mRNA-LNPs) without compromise on data density.
• Quartz 96-well microplate platform: Chemically inert, UV-transparent, and thermally stable; compatible with acidic, basic, and organic solvent-containing formulations.
• Rapid thermal profiling: Full 96-well plate scan completed in under 60 seconds per temperature step; typical full ramp (25–95 °C) executed in ≤90 minutes.
• Robust thermal control: Peltier-based heating/cooling with ±0.1 °C accuracy and <0.05 °C uniformity across all wells.
• Automated sealing integration: Compatible with heat-seal films and pressure-sensitive adhesives to prevent evaporation and cross-contamination during extended runs.

Sample Compatibility & Compliance

The Aunty system accommodates a broad range of biopharmaceutical samples—including monoclonal antibodies, Fc-fusion proteins, enzymes, virus-like particles (VLPs), adeno-associated virus (AAV) capsids, and lipid nanoparticle (LNP)-formulated therapeutics. Its quartz microplate design ensures compatibility with high-salt buffers, chaotropes (e.g., guanidine HCl), reducing agents (e.g., TCEP), and surfactants (e.g., polysorbate 20/80) without optical interference or surface adsorption artifacts. From a regulatory standpoint, the instrument’s firmware and accompanying software support 21 CFR Part 11 compliance through role-based user access, electronic signature capture, immutable audit trails, and secure data export (CSV, PDF, XML). It aligns with ICH Q5C (stability testing of biotechnological/biological products) and supports documentation required for IND, BLA, and MA submissions.

Software & Data Management

Aunty is operated via Unchained Labs’ proprietary Stabiliti™ Software Suite, a Windows-based application built for reproducible method definition, real-time visualization, and multivariate analysis. Users define custom thermal profiles, select wavelength bands for DSF deconvolution (e.g., 330 nm/350 nm ratio), set SLS integration times, and configure DLS correlation decay models—all within a single protocol. Raw data are stored in a structured SQLite database with embedded metadata (operator ID, timestamp, plate barcode, environmental logs). Batch analysis tools enable automated Tm extraction using derivative peak detection, Tagg assignment via SLS inflection point analysis, and kD/B22 calculation from second virial coefficient modeling. Exported reports include annotated plots, statistical summaries (mean ± SD, CV%), and pass/fail flags against user-defined stability thresholds.

Applications

• Early-stage formulation screening: Rapid identification of optimal pH, buffer species, excipients, and salt concentrations that maximize Tm and suppress aggregation.
• Biosimilar developability assessment: Comparative thermal fingerprinting against reference products to confirm structural equivalence.
• Process-related impurity impact studies: Evaluation of host cell proteins or leachables on product stability profiles.
• Viral vector stability monitoring: Quantification of capsid integrity loss (via Tm shift) and particle size distribution broadening (via DLS) under storage or freeze-thaw stress.
• Accelerated stability prediction: Correlation of short-term thermal denaturation metrics with long-term real-time stability data per ICH Q1A(R2).
• Forced degradation studies: Controlled exposure to oxidative, acidic, or alkaline stress followed by Aunty-based stability readout.

FAQ

Can Aunty measure stability of membrane proteins solubilized in detergents?
Yes—its quartz microplates tolerate common detergents (e.g., DDM, CHAPS, OG) without signal quenching or scattering artifacts, and DSF spectral deconvolution allows discrimination between protein unfolding and detergent micelle transitions.
Is calibration traceable to NIST standards?
Temperature calibration is performed using certified reference materials traceable to NIST SRM 1965 (protein thermal transition standard); optical path length and scattering angle geometries are factory-validated and documented in the instrument certificate of conformance.
Does Aunty support kinetic aggregation assays at isothermal conditions?
Yes—users may configure isothermal holds at any temperature (e.g., 40 °C for 24 h) with continuous SLS/DLS acquisition to monitor time-dependent aggregation without thermal ramping.
How does Aunty handle highly viscous or turbid samples?
SLS and DLS modules incorporate automatic gain adjustment and multi-angle scattering normalization; DSF employs background subtraction algorithms optimized for high-absorbance matrices, ensuring robust signal recovery up to OD₃₅₀ ≈ 1.2.
Can raw data be reprocessed with updated algorithms after acquisition?
Yes—Stabiliti™ supports offline reanalysis of archived .stb files using current or legacy algorithm versions, preserving full traceability and enabling retrospective method updates without re-running experiments.