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| Determination of Protein Content of Alhydrogel-absorbed Vaccine Using O-phthalaldehyde (OPA) Fluorometric Method |
| LEI Ning1, ZHANG Yuan2, XIAO Hai-bo2, ZENG Qing-le1 |
1. College of Material and Chemistry & Chemical Engineering, Chengdu University of Technology, Chengdu 610059, China;
2. Chengdu Fronter BioScience Inc., Chengdu 610041, China |
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Abstract Objective: To develop a generic method for the determination of protein content in alhydrogel (aluminum) absorbed vaccine formulation using o-phthalaldehyde (OPA). Method: based on the theory that OPA can react with amino acids under alkaline conditions and give strong fluorescence, in the presence of 2-mercaptoethanol, a fluorescent method without antigen extraction was developed and the specificity, linearity range, accuracy and precision were evaluated. The integrity of the vaccine formulation was also investigated by SDS-PAGE. Results: a good linearity was obtained from 0.02mg/ml to 0.16mg/ml of antigen content, the accuracy of 90%~115% and CV% less than 15% were achieved. Little protein disassociation was also observed after CpG was added into the formulation. Conclusion: the OPA fluorescent method was simple and can be further used not only in the laboratory but also in routine quality control of the vaccine formulation with alhydrogel.
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Received: 21 December 2010
Published: 25 July 2011
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