摘要
目的寻找分离纯化牛视网膜Ｓ抗原的快速高效方法。方法选用ＤＥＡＥＳｅｐｈａｒｏｓｅＦａｓｔＦｌｏｗ（ＤＥＡＥＳｅｐｈａｒｏｓｅＦＦ）凝胶及自动层析系统一步层析快速提纯Ｓ抗原；聚丙烯酰胺凝胶电泳确定分子量；等电聚焦（ｉｓｏｅｌｅｃｔｒｉｃｆｏｃｕｓｉｎｇ，ＩＥＦ）电泳确定等电点；高效液相色谱（ｈｉｇｈｐｅｒｆｏｒｍａｎｃｅｌｉｑｕｉｄｃｈｒｏｍａｔｏｇｒａｐｈｙ，ＨＰＬＣ）确定纯度。结果Ｓ抗原波峰在ＮａＣｌ浓度为０．１４～０．２ｍｏｌ／Ｌ时被洗脱，每个视网膜能提纯３００～４００μｇＳ抗原。纯化Ｓ抗原的分子量为５５ｋＤａ，等电点为６．５５，ＨＰＬＣ分析显示单一波峰，保留时间为１１．５６ｍｉｎ。
ObjectiveTo find a rapid and effective method for isolation of retinal S antigen.MethodsS antigen was isolated from bovine retina by ion exchange chromatography on DEAE Sepharose Fast Flow(DEAE Sepharose FF)and automatic Gradi Frac System.The isolated S antigen was analysed with sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS PAGE),isoelectric focusing(IEF)and high performance liquid chromatogrphy(HPLC).ResultsS antigen peak was eluted in 0.14～0.2mol/L NaCl linear gradient elution.Each retina resulted in 300～400 μg of purified S antigen.The analytical studies showed that the molecular weight of the bovine purilfied S antigen was 55 kDa,its isoelectric point was 6.55.HPLC showed a single peak.Its retention time was 11.56 min.ConclusionThis method simply and rapidly yields a highly purified S antigen.
出处
《眼科研究》
CAS
CSCD
1998年第2期96-98,共3页
Chinese Ophthalmic Research
基金
国家自然科学基金
霍英东教育基金会
广东省自然科学基金
