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Assessment of the Protein–Protein Interactions in a Highly Concentrated Antibody Solution by Using Raman Spectroscopy

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ABSTRACT

Purpose

To investigate the protein–protein interactions of a highly concentrated antibody solution that could cause oligomerization or aggregation and to develop a better understanding of the optimization of drug formulations.

Methods

In this study, we used Raman spectroscopy to investigate the structure and interactions of a highly concentrated antibody solution over a wide range of concentrations (10–200 mg/mL) with the aid of a multivariate analysis.

Results

Our analysis of the amide I band, I 856 /I 830 of Tyr, and the relative intensity at 1004 cm−1 of the Phe and OH stretching region at around 3000 cm−1 showed that across this wide range of concentrations, the secondary structure of the IgG molecules did not change; however, short-range attractive interactions around the Tyr and Phe residues occurred as the distance between the IgG molecules decreased with increasing concentration. Analysis of the OH stretching region at around 3000 cm−1 showed that these short-range attractive interactions correlated with the amount of hydrated water around the IgG molecules.

Conclusions

Our data show that Raman spectroscopy can provide valuable information of the protein–protein interactions based on conformational approaches to support conventional colloidal approaches, especially for analyses of highly concentrated solutions.

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Abbreviations

MCR-ALS:

Multivariate curve resolution-alternating least squares

PCA:

Principal components analysis

REV:

Reduced-eigenvalue

RMSECV:

Root-mean-square prediction error of cross-validation

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ACKNOWLEDGMENTS AND DISCLOSURES

We thank our colleague Dr. Daisuke Irikura (Horiba, Ltd.) for helpful comments on the results and discussion.

Author information

Authors and Affiliations

  1. Advanced R&D Center, Horiba, Ltd., 2 Miyanohigashi, Kisshoin, Minami-ku, Kyoto, 601-8510, Japan

    Chikashi Ota & Shintaro Noguchi

  2. Department of Bioengineering, School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Satoru Nagatoishi & Kouhei Tsumoto

  3. Laboratory of Medical Proteomics, Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, Japan

    Kouhei Tsumoto

  4. Drug Discovery Initiative, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Kouhei Tsumoto

Authors
  1. Chikashi Ota
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  2. Shintaro Noguchi
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  3. Satoru Nagatoishi
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  4. Kouhei Tsumoto
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Corresponding author

Correspondence to Chikashi Ota.

Electronic supplementary material

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ESM 1

The results of PCA (REV, RMSECV) are supplied as Supporting Information 1,3,5 (Fig. S1,3,5). Concentration dependence of Raman spectra in CH and OH stretching region (Fig. S2), the abstract spectra of MCR-ALS analysis (Fig. S4) and distance between the molecules of the IgG solution as a function of concentration (Fig. S6) are also supplied as Supporting Information 2,4,6. (DOCX 940 kb)

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Ota, C., Noguchi, S., Nagatoishi, S. et al. Assessment of the Protein–Protein Interactions in a Highly Concentrated Antibody Solution by Using Raman Spectroscopy. Pharm Res 33, 956–969 (2016). https://doi.org/10.1007/s11095-015-1842-8

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  • DOI: https://doi.org/10.1007/s11095-015-1842-8

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