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Mu L.,Chengdu Rongsheng Pharmaceutical Co. | Lu T.,Chengdu Rongsheng Pharmaceutical Co. | Wang Q.-C.,Chengdu Rongsheng Pharmaceutical Co. | Li W.,Chengdu Rongsheng Pharmaceutical Co. | Yu W.,Chengdu Rongsheng Pharmaceutical Co.
Chinese Journal of Biologicals | Year: 2015

Objective To develop a method for preparation of human coagulation factor VIII (FVIII) by ion exchange chromatography as well as the formula of stabilizer. Methods The plasma-derived cryoprecipitate was purified by PEG precipitation plus ion exchange chromatography, in which the virus was inactivated by S/D and dry heating at 80°C for 72 h. The inactivation effect was verified, and the optimal formula of stabilizer was developed. Results The purity of FVIII increased by about 110 folds after PEG precipitation, while by more than 30 folds after ion exchange chromatography. Both S/D method and dry heating at 80°C for 72 h showed satisfactory inactivation effects. The optimal formula of stabilizer consisted of 0.01 mol/L sodium citrate, 0.001 mol/L calcium chloride, 8 mg/ml albumin and 40 mg/ml arginine hydrochloride. Conclusion The developed method may be used for preparation of FVIII with high purity, stability and safety. The screened formula of stabilizer was resistant to inactivation by dry heating. Source


Nie Y.-T.,Chengdu Rongsheng Pharmaceutical Co. | He T.-P.,Chengdu Rongsheng Pharmaceutical Co. | Lei T.,Chengdu Rongsheng Pharmaceutical Co. | Ge Y.-H.,Chengdu Rongsheng Pharmaceutical Co. | And 6 more authors.
Chinese Journal of Biologicals | Year: 2010

Objective: To construct an immune antibody library and screen sub-nM affinity human antibody. Methods: The genes encoding variable region were amplified from the high titer human B lymphocytes immunized with tetanus toxoid by PCR to construct an immune antibody library, from which high affinity human antibody was screened. The obtained antibody gene fragments were transferred into whole molecules which were expressed and purified then compared with human polyclonal antibody against TT. Results: An immune phage antibody library against human tetanus, with a capacity of 4. 5 × 106, was constructed, of which the diversity met the requirements for design of primers. The human antibody with a relative affinity of 10-10 M was obtained, of which the neutralizing activity was higher than that of human polyclonal antibody. Conclusions: Sub-nM affinity human antibody was obtained by the construction of immune antibody library with small capacity by designing primers according to the frequency of antibody gene. The strategy may be used for the development of human antibody against infectious diseases. Source

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