TitrationAnalysis : a tool for high throughput binding kinetics data analysis for multiple label-free platforms

• Published in: Gates open research Vol. 7; p. 107
• Main Authors: Li, Kan, Huntwork, Richard H C, Horn, Gillian Q, Alam, S Munir, Tomaras, Georgia D, Dennison, S Moses
• Format: Journal Article
• Language: English
• Published: United States F1000 Research Ltd 2023
• Subjects: antibody binding; Biolayer Interferometry; eng; high-throughput kinetics analysis; non-linear curve fitting; Surface Plasmon Resonance; Surface Plasmon Resonance; non-linear curve fitting; high-throughput kinetics analysis; Biolayer Interferometry; antibody binding
• ISSN: 2572-4754; 2572-4754
• DOI: 10.12688/gatesopenres.14743.1

Label-free techniques including Surface Plasmon Resonance (SPR) and Biolayer Interferometry (BLI) are biophysical tools widely used to collect binding kinetics data of bimolecular interactions. To efficiently analyze SPR and BLI binding kinetics data, we have built a new high throughput analysis tool named the
. It can be used as a package in the Mathematica scripting environment and ultilize the non-linear curve-fitting module of Mathematica for its core function. This tool can fit the binding time course data and estimate association and dissociation rate constants (
and
respectively) for determining apparent dissociation constant (
) values. The high throughput fitting process is automatic, requires minimal knowledge on Mathematica scripting and can be applied to data from multiple label-free platforms. We demonstrate that the
is optimal to analyze antibody-antigen binding data acquired on Biacore T200 (SPR), Carterra LSA (SPR imaging) and ForteBio Octet Red384 (BLI) platforms. The
,
and
values derived using
very closely matched the results from the commercial analysis software provided specifically for these instruments. Additionally, the
tool generates user-directed customizable results output that can be readily used in downstream Data Quality Control associated with Good Clinical Laboratory Practice operations. With the versatility in source of data input source and options of analysis result output, the
high throughput analysis tool offers investigators a powerful alternative in biomolecular interaction characterization.