New method to produce equine antirabies immunoglobulin F(ab ')(2) fragments from crude plasma in high quality and yield

作者：Kittipongwarakarn Sukanda; Hawe Andrea; Tantipolphan Ruedee****; Limsuwund Kornvika; Khomvilai Sumana; Puttipipatkhachorn Satit^*; Jiskoot Wim

来源：European Journal of Pharmaceutics and Biopharmaceutics, 2011, 78(2): 189-195.

DOI：10.1016/j.ejpb.2011.02.018

• Antirabies serum
• Equine antirabies immunoglobulin
• Purification
• Pepsin digestion
• Protein G affinity chromatography
• Protein A affinity chromatography
• F(ab ')(2) fragments

Rabies is still a major cause of human deaths in several developing countries. According to the World Health Organization, administration of antirabies serum or antirabies immunoglobulin is recommended for patients who have experienced a category-III exposure to rabies. Improvement of antirabies immunoglobulin production is required to enhance safety and efficacy of the products. In this paper, a new method to produce equine antirabies immunoglobulin F(ab')(2) fragments from crude plasma is proposed. First, protein G affinity chromatography was used to purify IgG from equine plasma. Moreover, purification of IgG was shown to facilitate its digestion by pepsin. Compared to the direct digestion of crude plasma, a lower amount of pepsin and a shorter digestion time were required to completely digest the purified IgG to F(ab')(2). Complete digestion of purified IgG to F(ab')(2) was achieved at a pepsin/IgG (w/w) ratio of 5:45 with preservation of structure and potency. Finally, purification of F(ab')2 was accomplished by a combination of protein A affinity chromatography and ultrafiltration with a 50-kDa nominal molecular weight cut-off membrane. The new process resulted in 68.9 +/- 0.6 (%) total recovery of F(ab')(2) and a F(ab')(2) product of high potency.

• 出版日期2011-6