Spectra Manager BeStSel

Secondary structure information using BeStSel algorithm (left) Automatic Y-axis conversion and concentration calculation using amino acid sequence and absorption (right)

THERMAL DENATURATION OF ANTIBODIES

Analyzed with BeStSel

Application: Secondary Structure Prediction of Therapeutic Antibodies

Here, we compare Herceptin®, a well known breast cancer drug, and human IgG. The structure of the antibody at various temperatures was monitored by measuring the secondary structure CD spectrum from 30° to 85°C.

BeStSel offers fractional analysis of eight secondary structure elements, including three types of antiparallel β-sheet (Left-twisted, Right-twisted, Relaxed). Significant differences between Herceptin® and human-IgG in relaxed β-sheet change may reflect the formation of human IgG aggregates rather than simple denaturation as seen for Herceptin. BeStSel proves to be a powerful tool as this difference is clearly seen with the β-sheet model.

CD spectra of Herceptin® (left) and CD Spectra of Human IgG (right)
Fractions of antibody secondary structure as a function of temperature.

 

Spectra Manager BeStSel utilizes the ‘BeStSel engine’, an algorithm developed by Dr. József Kardos and Dr. András Micsonai, et. al. at ELTE (Eötvös Loránd University, Hungary).