boltzmann sigmoidal fitted curves  (GraphPad Software Inc)

Journal: Communications Biology

Article Title: Stability convergence in natural antibodies with ultra-long hypervariable loops

doi: 10.1038/s42003-025-08036-5

Figure Lengend Snippet: a Structure of a bovine Fab NC-Cow1 (PDB:6OO0) with an ultra-long CDR-H3 (gray color). Schematic figure and ulCDR sequence alignment of the Fab variants in this work. b Non-reducing SDS-PAGE of cell supernatants with secreted Fab fragments and a purified NC-Cow1 wt reference. c Chromatograms and molecular mass in SEC-MALS. d Mean values ( n = 2) of molecular masses calculated from MALS. e Exemplary melting curves in SUPR-DSF. f Apparent onset temperatures of unfolding T on and melting temperatures T m obtained from SUPR-DSF measurements. Mean values ( n = 3). g Slope factors of fitted Boltzmann functions obtained from SUPR-DSF melting curves. A smaller slope factor indicates a steeper transition (more cooperative unfolding). Mean values ( n = 3; SD shown in Table ). h Apparent hydrodynamic radii R h from DLS. Mean values ( n = 3) with SD. i Apparent hydrodynamic radii R h during heating ramp with DLS. j Aggregation onset temperatures T agg obtained from heat-ramped DLS. Mean values ( n = 3) with SD.

Article Snippet: For comparing the slopes of the melting curves, a sigmoidal Boltzmann fit was performed using GraphPad Prism version 8.0.1 for Windows, GraphPad Software, Boston, Massachusetts USA, www.graphpad.com .

Techniques: Sequencing, SDS Page, Purification