目的探索包裹破伤风类毒素蛋白（TT）的聚乳酸(PLA）及聚乳酸/聚乙醇酸(PLG)共聚微球的制备及其体外释放特性。方法采用溶剂蒸发技术,先以一种小分子量生物染料伊文思蓝为包裹物模型，通过显微镜观察和比色分析探索微球的制备特性和最佳条件。在此基础上制备包裹TT的PLA和 PLG微球。结果微球的制备过程中存在着内水相蛋白朝外水相的流失，当聚合物浓度加大到25%～30%时可使蛋白流失大大减少。包裹蛋白在微球中的分布与微球粒径的重量分布成正相关关系；随着微球粒径的增大，微球所载蛋白的体外释放高峰期延迟。结论通过本模型条件可制备外观高质量的微球，并可通过制备不同粒径大小的微球来控制包裹蛋白在体外和体内的释放高峰期。
Abstract
OBJECTIVE To investigate the preparation and release properties of poly lactid acid(PLA) and poly lactidco glycolide acid (PLG) microspheres (MS) containing tetanus toxoid (TT) protein.METHOD The preparation property and optimum techniques of PLAMS were investigated by microscope and colorimetry using Even's blue, a low molecular weight bio dye as a model encapulating material.According to the optimum techniques,the microspheres containing TT protein were prepared.RESULT During the preparation of microspheres containing TT protein,the loss of TT protein from internal aqueous phase to external aqueous phase occurred and the loss of TT decreased when the concentration of polymer in dichlormethane increased to 25%～30%.The distribution of encapulated TT protein in microspheres was relevant with the weight distribution of different size of microspheres.In vitro,the peak release time of encapulated TT protein was delayed when the diameter of microspheres increased. CONCLUSION The TT proteinloading PLA and PLG microspheres with a smooth and nonporous surface was prepared by optimum techniques.The different size of microspheres containing TT protein can also be prepared for the need of controlled peak release time of encapulated TT protein in vitro and in vivo.
关键词
聚合物微球 /
破伤风类毒素蛋白 /
特性
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Key words
polymer microspheres /
tetanus toxoid protein /
properties
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参考文献
[1]Haeckel S,Sachse A,Albayrak C，et al. Preparation and characterization of insulin-loaded microcapsules produced by the induced phase separation method[A].In: Preceeding of the second world meeting [C]. APGI/APV Paris，1998:196～200.
[2]Ciftci K,Kas HS,Hincal AA，et al.In vitro and in vivo evaluation of PLAGA(50/50) microspheres containing 5-fluorouracil prepared by a solvent evaporation method[J]. Int J Pharm, 1996，131:73～82.
[3]Cheng A,Gupta RK.Stabilization of tetanus toxoid in poly(DL-lactic-co-glycolic acid) microspheres for the controlled release of antigen[J]. J Pharm Sci, 1996, 85(2):129～132.
[4]Kersten GFA,Gander B.Biodegradable microspheres as vehicles for antigens[A].In:Kaufmann SHE ed. Concepts in Vaccine Development [M].Berlin,New York:Walter de Gruyter & Co,1996:265～302.
[5]Sharif S,O'Hagan DT.A comparison of alternative methods for the determination of the levels of proteins entrapped in (lactide-co-glycolide) microparticles[J]. Inter J Pharm, 1995,115:259～263.
[6]Singh M,Li XM,Mcgee JP, et al. Controlled releases microparticles as single dose hepatitis B vaccine: evaluation of immunogenicity in mice[J]. Vaccine, 1997,15(5):475～481.
[7]Smith PK,Krohn RI,Hermanson GT. et al. Measurement of protein using bicinchoninic acid[J]. Anal Biochem, 1985,150:76～85.
[8]Eldridge JH,Steas J,Meulbroek JA，et al. Biodegradable and biocompatible poly(DL-lactide-co-glycolide) microspheres as an adjuvant for staphylococcal enterotoxin B toxoid which enhances the level of toxin-neutralizing antibodies[J]. Infect Immu, 1991,9:2978～2986.
[9]Jeffery H,Davis SS,O′Hagen DT.The preparation and characterization of poly(lactide-co-glycolide) microspheres.Ⅱ.The entrapment of a model protein using a (water-in-oil)-in-water emulsion by solvent evaporation technique[J]. Pharm Res, 1993,10(3):262～268.
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