Orthogonal Assessment of Antibody Secondary Structure
The JASCO qHOS spectral comparison program is capable of evaluating secondary structure similarity of proteins via CD and FTIR.
Quick and quantitative orthogonal assessment of protein secondary structure with FTIR and CD.
Similarity Assessment for Secondary Structure of Biosimilars
The far-UV/CD and FTIR spectra of the biosimilar RIABNITM are in excellent agreement with those of the innovator MabThera®, and the distributions of the distances between MabThera® and RIABNITM are close to each other. The p-value is larger than the significance level of 0.05.
Orthogonal Assessment of Antibody Tertiary Structure
With JASCO qHOS, Raman and CD spectroscopy comes together to quantitatively determine spectral similarity for monoclonal antibodies.
Evaluation of Tertiary structure is quickly performed using the qHOS statistical package using a known reference and sample data.
Similarity Assessment for Tertiary Structure of Biosimilars
Similar to the secondary structure, the tertiary structure of MabThera® and RIABNITM show excellent agreement in the near-UV/CD and Raman spectra (Figs. 3a and 3b), and the distribution of the distances between MabThera® and RIABNITM are close to each other (Figs. 3c and 3d). The p-value is larger than the significance level of 0.05.
qHOS provides similarity evaluation with flexibility of methods. Our selected weighting methods avoid issues with spectral intensity weighting often misused with CD.
| Three Different Statistical Tests | Two Weighting Methods | |||
|---|---|---|---|---|
| Student t-test | For analyzing difference a single sample spectrum | Noise | Gives more weight to spectral regions with less noise via light level | |
| Welch's t-test | For analyzing differences for multiple samples | Spectra Change | Weights region where the spectrum is known to change using a given native and denatured spectrum | |
| Two one sided t-test (TOST) | Establishes sample equivalence through two t-tests |
Click below to view our PEGS 2023 poster on secondary structure.