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Determination of Adsorbed Protein Concentration in Aluminum Hydroxide Suspensions by Near-Infrared Transmittance Spectroscopy

Volume 62, Number 7 (July 2008) Page 784-790

Lai, Xuxin; Zheng, Yiwu; Jacobsen, Susanne; Larsen, Jørgen N.; Ipsen, Henrik; Løwenstein, Henning; Søndergaard, I.B.


Analysis of aluminum hydroxide based vaccines is difficult after antigen adsorption. Adsorbed protein is often assessed by measuring residual unadsorbed protein for quality control. A new method for the direct determination of adsorbed protein concentration in suspension using near-infrared (NIR) transmittance spectroscopy is proposed here. A simple adsorption system using albumin from bovine serum (BSA) and aluminum hydroxide as a model system is employed. The results show that the NIR absorbance at 700-1300 nm is correlated to the adsorbed BSA concentration, measured by the ultraviolet (UV) method, using the partial least square regression (PLSR) method to construct a calibration model. The linear concentration range of adsorbed BSA is from 0 to 1.75 mg/mL by using 10 mm path length cuvettes. The influence of the sedimentation in suspension, different buffers, and different aluminum hydroxide batches was investigated in this study. It shows that the batch variation is the main influence factor of this method, while the buffer variation has no influence. However, the pretreatment of spectral data by subtracting spectra of BSA blank control (aluminum hydroxide without BSA) can significantly reduce the batch influence, and the NIR predicted results show good agreement with the reference values. The NIR method might be the only direct method for the determination of adsorbed protein concentration in suspension so far. It is a nondestructive method, and it has great advantage for use in vaccine production as a method for quality control and quality assurance.