DIY in Lunatic and Stunner with the Homebrew toolkit

Importance of the Topic

The accurate and high-throughput quantification and characterization of biologics such as proteins, antibodies, and nucleic acids is vital in research, quality control, and biopharmaceutical development.
Traditional workflows often involve manual dilutions, multiple instruments, and complex data processing, leading to extended turnaround times and potential errors.
Integrating micro-volume UV/Vis analysis with dynamic light scattering within a unified platform addresses these challenges by enabling rapid concentration measurements, aggregation assessment, and viscosity determination using minimal sample volumes.

Objectives and Overview

This application note demonstrates the capabilities of the Lunatic micro-volume UV/Vis reader and the Stunner combined UV/Vis and DLS instrument equipped with the Homebrew toolkit.
Custom applications can streamline sample processing by automating background correction, statistical analysis, and viscosity or aggregation measurements.
Two case studies are presented: protein quantification with replicate statistics on Lunatic, and DLS-based viscosity measurements on Stunner.

Methodology

Protein samples of bovine IgG at 1 and 10 mg/mL in PBS were measured in triplicate using 2 µL per well in High Lunatic plates.
Homebrew protein applications applied turbidity background correction and an E1% of 13.7 to determine concentrations, averages, standard deviations, and coefficients of variation.
For viscosity measurements, 100 nm NIST standard beads were added to glycerol solutions of varying concentrations and to monoclonal antibody formulations in histidine–arginine buffer.
Diffusion coefficients were obtained from four DLS acquisitions of 5 seconds each, and solution viscosities were calculated via the Einstein–Stokes equation.

Instrumentation

• Lunatic UV/Vis micro-volume spectrophotometer: measures full absorbance spectra from 0.02–200 mg/mL with 2 µL sample volumes and microfluidic 96-well plates to prevent cross-contamination.
• Stunner combined UV/Vis and dynamic light scattering instrument: acquires simultaneous concentration and size measurements for up to 96 samples in one hour using 2 µL volumes.
• Homebrew toolkit within Lunatic & Stunner Client v6.0: provides custom workflows, multiple background correction options (turbidity, intelligent single-point, user-defined, average), and built-in statistical and arithmetic functions for creating tailored outputs.

Main Results and Discussion

Replicate measurements of 10 mg/mL IgG yielded consistent concentration values with low coefficients of variation, illustrating robust statistical analysis within Homebrew.
Viscosities of glycerol solutions determined by Stunner closely matched published CRC Handbook values, validating the DLS-based approach.
In monoclonal antibody formulations, Stunner-derived viscosities agreed closely with those measured by a RheoSense µVISC viscometer, demonstrating the accuracy of the micro-volume DLS method.

Contributions and Practical Applications

Homebrew reduces hands-on time and sample consumption by integrating data acquisition, custom analysis, and statistics into a single interface.
The combined UV/Vis and DLS platform enables early detection of aggregation and viscosity changes, supporting formulation development and quality control in biopharmaceutical workflows.

Future Trends and Potential Applications

Expanding Homebrew applications to additional biomolecule classes and advanced spectral analyses can further streamline high-throughput workflows.
Integration with laboratory automation systems and informatics platforms will enhance data management and reproducibility.
Emerging algorithms for pattern recognition and machine learning could be incorporated to provide predictive insights into sample behavior.

Conclusion

Lunatic and Stunner, augmented by the Homebrew toolkit, offer versatile, high-throughput solutions for quantitative and qualitative analysis of biologics using minimal sample volumes.
The combined capabilities of micro-volume UV/Vis and DLS, along with customizable background correction and statistical functions, deliver accurate, rapid results for research and QC applications.

References

1. Connolly B et al. Biophysical Journal. 2012;103(1):69–78.
2. Unchained Labs. Spot aggregation early with B22 and kD on Stunner. Application Note. 2019.
3. Haynes W, editor. CRC Handbook of Chemistry and Physics. 95th ed. CRC Press; 2014.