Volume 101, Issue 7, Pages (October 2011)

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Volume 101, Issue 7, Pages 1749-1757 (October 2011) High-Throughput Analysis of Concentration-Dependent Antibody Self-Association Shantanu V. Sule, Muppalla Sukumar, William F. Weiss, Anna Marie Marcelino-Cruz, Tyler Sample, Peter M. Tessier Biophysical Journal Volume 101, Issue 7, Pages 1749-1757 (October 2011) DOI: 10.1016/j.bpj.2011.08.036 Copyright © 2011 Biophysical Society Terms and Conditions

Figure 1 Immobilization of mAbs on gold nanoparticles. (A) UV-visible absorbance spectra of gold particles (black circles), gold-mAb1 conjugates (gray circles), and gold-mAb2 conjugates (white triangles). (B) DLS analysis of gold particles, antibodies (wild-type and thiolated), and gold-antibody conjugates. Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions

Figure 2 Analysis of variable antibody self-association at near-neutral pH. (A) Plasmon wavelengths (λp) of mAb1 and mAb2 conjugates at pH 6.5 and 6, respectively. (B) SLS measurements of WAMW values for mAb1 and mAb2 at 6 mg/mL. Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions

Figure 3 SINS analysis of antibody self-association as a function of pH and ionic strength. Plasmon wavelengths (λp) of (A) gold-mAb1 and (B) gold-mAb2 conjugates at pH 4.3 (solid circles) and pH 6.5/6 (open triangles). Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions

Figure 4 SLS analysis of antibody self-association as a function of pH and ionic strength. WAMW measurements for (A) mAb1 and (B) mAb2 (6 mg/mL) at pH 4.3 (solid circles) and pH 6.5 (open triangles). Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions

Figure 5 Correlation between SINS and SLS measurements of antibody self-association. Plasmon wavelengths (λp) for gold-antibody conjugates plotted versus WAMW values obtained at antibody concentrations of (A) 6 and (B) 42 mg/mL. Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions

Figure 6 Comparison of self-association behavior for three mAbs. Measurements of (A) WAMW (6 mg/mL) and (B) plasmon wavelengths (λp) at pH 6.5 (mAb1 and mAb3) and pH 6 (mAb2). The salt concentration was 150 mM NaCl. Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions

Figure 7 Self-association behavior of a polyclonal antibody as a function of pH and ionic strength. Plasmon wavelengths (λp) of polyclonal antibody-gold conjugates at pH 4.3 (black circles), pH 6 (gray squares) and pH 6.5 (white triangles). Biophysical Journal 2011 101, 1749-1757DOI: (10.1016/j.bpj.2011.08.036) Copyright © 2011 Biophysical Society Terms and Conditions