Obtaining recombinant chaperon CroEL and its immunological cross-reactivity with Hsp60

Abstract

The method of obtaining and purification of recombinant chaperon GroEL (prokaryotic homolog of eukaryotic chaperon Hsp60) including the protein expression in E. coli cells, precipitation with saturated ammonium sulphate from a producent lysate, gel-filtration on Sephacryl-300 column and ion-exchange chromatography on MonoQ HR 5/5 column, is described. The present method allows effective obtaining of the preparative amount of the GroEL protein of 95 % purity. The recombinant protein was used for the anti-GroEL antibodies production and synthesis of the affine column. The immunological cross-reactivity of polyclonal affinepurified anti-GroEL antibodies and members of Hsp60 family from different organs and cells lysates of different mammals, from mouse to human, have been revealed, which allows using these antibodies for research of Hsp60 expression alterations and cell localization at human autoimmune and cancer pathologies. © L.N. Kapustian, R.G. Kyyamova, V.S. Gryshkova, A.G. Terentiev, L.L. Sidorik.