Liu Y.-X.,Chifeng Boen Pharmaceutical Biology Co. |
Zhao Z.-P.,Chifeng Boen Pharmaceutical Biology Co. |
Luo D.-Y.,Chifeng Boen Pharmaceutical Biology Co. |
Chen Z.-W.,Chifeng Boen Pharmaceutical Biology Co. |
And 3 more authors.
Chinese Journal of Biologicals | Year: 2011
Objective: To develop a novel procedure for preparation of equine immunoglobulin F(ab′) 2 against tetanus toxin. Methods: Tetanus toxoid (TT) and recombinant tetanus toxin C fragment (rTT-C) were prepared and purified, then mixed at a ratio of 2:1. Horse were immunized with the mixture TT/rTT-C and, when the serum antibody titer reached 3 500 IU/ml, the sera were collected and inactivated, from which the foreign protein was removed, then digested with pepsin. The prepared F(ab′) 2 was purified by DEAE column chromatography, and analyzed for neutralizing efficacy, safety and stability. Results: The concentrations of purified TT and rTT-C were 70 000 Lf/L and 4-5 mg/L, while the purities were more than 95% and more than 96%, respectively. The recovery rate, purity and neutralizing antibody titer of purified TAT- F (ab′) 2 were 1.4%, more than 91% and more than 15 000 IU/ml respectively, while the safety and stability met the requirements in Chinese Pharmacopeia (Volume III, 2005 edition). The validity period was determined as 18 months temperately. Conclusion: A novel procedure for preparation of equine immunoglobulin F(ab′) 2 against tetanus toxin was developed, and the quality of product was equal to or even higher than that of imported product, which provided a technical support for upgrading of equine antiserum of the same kind.