DIY in Lunatic and Stunner with the Homebrew toolkit

Importance of the topic

In biopharmaceutical research and quality control, rapid and accurate protein quantification and characterization are critical to accelerate development timelines, ensure product consistency, and reduce sample consumption. Microvolume UV/Vis analysis and dynamic light scattering (DLS) methods can streamline workflows by minimizing manual dilutions and automating data processing. The Unchained Labs Lunatic and Stunner platforms, enhanced with the Homebrew toolkit, offer a flexible environment to design custom assays that combine high-throughput spectral analysis with statistical evaluation and viscosity measurement, addressing the growing demand for integrated analytical solutions in life sciences.

Objectives and article overview

This application note demonstrates how to employ the Homebrew toolkit on Lunatic and Stunner instruments to:

• Develop custom UV/Vis-based protein quantification applications with built-in statistical analysis.
• Implement flexible background correction strategies to optimize data accuracy.
• Integrate DLS-based measurements of solution viscosity and protein aggregation propensity.
• Streamline workflows for high-throughput assays using minimal sample volumes.

The note provides examples of setting up replicate analysis for bovine IgG quantification and using NIST bead standards to calculate buffer viscosity via the Einstein–Stokes relationship.

Instrumentation

The following equipment and software are described:

• Lunatic microvolume UV/Vis spectrophotometer with dynamic range from 0.02 to 200 mg/mL and 2 µL sample volume capacity.
• Stunner instrument combining UV/Vis spectroscopy and dynamic light scattering (DLS) for simultaneous concentration and size determination of proteins.
• Homebrew software toolkit (Client v6.0) for designing custom data processing workflows, background correction methods, and output variables.
• High Lunatic and Stunner microplate formats compatible with 96-sample throughput and automation integration.
• NIST standard 100 nm bead solutions for viscosity calibration.

Methodology

Protein quantification workflow:

• Prepare bovine IgG samples at 1 mg/mL and 10 mg/mL in PBS, load 2 µL in triplicate into Lunatic plates, with PBS as blank.
• Collect full UV/Vis absorbance spectra and apply turbidity correction or user-defined background adjustments.
• Calculate concentration using absorbance at 280 nm and an input E1% extinction coefficient of 13.7.
• Utilize Homebrew functions to compute average, standard deviation, and coefficient of variation for replicates sharing sample names.

DLS-based viscosity measurement:

• Prepare glycerol series in PBS and monoclonal antibody solutions in histidine buffer; add 100 nm NIST beads.
• Load 2 µL into Stunner plates and perform multiple DLS acquisitions to determine diffusion coefficients.
• Apply the Einstein–Stokes equation rearrangement to calculate viscosity from diffusion data, temperature, Boltzmann constant, and bead radius.
• Compare viscosity readings with standard viscometer measurements for validation.

Main findings and discussion

Protein quantification:

• Homebrew-generated protein assays on Lunatic yielded reproducible concentration measurements with low coefficient of variation across triplicates.
• Automated background correction options ensured accurate baseline adjustment without manual intervention.

Viscosity determination:

• DLS-derived viscosities for glycerol standards closely matched literature and reference viscometer values.
• Stunner successfully distinguished antibody monomer peaks from NIST beads, enabling reliable viscosity calculation for high-concentration formulations.

Together, these results illustrate how the combined UV/Vis and DLS capabilities, empowered by Homebrew, facilitate comprehensive sample characterization in a single platform.

Benefits and practical applications

The integrated Lunatic–Stunner workflow with Homebrew offers:

• High-throughput protein quantification without dilution steps, saving time and sample material.
• Customizable data processing and statistical outputs directly within the user interface.
• Versatile background correction methods tailored to diverse sample matrices.
• Rapid viscosity and aggregation propensity assessment for formulation development.
• Compatibility with automation systems and regulatory compliance (e.g., 21 CFR Part 11).

These advantages make the platform suitable for academic research, biopharmaceutical development, and routine quality control laboratories.

Future trends and possibilities

Emerging directions to expand the utility of the Homebrew-enabled Lunatic and Stunner systems include:

• Integration with machine learning algorithms for predictive analytics of protein stability and aggregation.
• Development of additional plug-in modules for nucleic acid quantification, lipid interactions, and nanoparticle characterization.
• Coupling with online process analytical technology (PAT) for real-time monitoring in bioprocessing.
• Enhanced miniaturization and multiplexing to increase throughput and reduce reagent consumption further.

Continued innovation in software and hardware could establish these platforms as central hubs for multi-parameter biomolecular analysis.

Conclusion

The Unchained Labs Lunatic and Stunner instruments, augmented by the Homebrew toolkit, deliver a powerful and flexible framework for integrated protein analysis. By merging microvolume UV/Vis quantification with DLS-based viscosity and size measurements, users can design tailored assays that improve throughput, data accuracy, and resource efficiency. The customizable Homebrew environment simplifies statistical reporting and background correction, enabling laboratories to adapt quickly to evolving analytical challenges in biopharmaceutical research and quality control.

References

1. Connolly B, et al. Weak interactions govern the viscosity of concentrated antibody solutions: High-throughput analysis using the diffusion interaction parameter, Biophysical Journal, 2012;103(1):69–78.
2. Unchained Labs. Spot aggregation early with B₂₂ and k_D on Stunner, Application Note, 2019.
3. Haynes W, ed. CRC Handbook of Chemistry and Physics, 95th ed., CRC Press, 2014.