[High-efficient renaturation of immobilized recombinant C-terminal fragment of human alpha-fetoprotein].
C-terminal fragment of a human oncofetal alpha-fetoprotein (AFP) may be used in targeted cytostatics delivery to malignant cells of many tumors. AFP fragment (from 404 to 595 amino acids residues of a full-sized protein) was cloned and produced in Escherichia coli cells, BL21 strain (DE3) in the form of inclusion bodies. To obtain a functionally active protein, is it necessary to renature the protein. The renaturation procedure of the AFP third domain (rAFP3D) is considerably complicated by the fact that the protein is hydrophobic and contains a large number of S-S bonds. A renaturation technique of rAFP3D immobilized on silicic metal chelate resin has been developed. The yield of renatured C-terminal fragment was no less than 60% with purity on the order of 98%. The developed technique has been applied for the first time for hydrophobic protein with a large number of S-S bonds. The approach can be applied for efficient renaturation of other hydrophobic proteins with a large number of disulfide bonds for scientific and practical purposes.
O A
OA
Sharapova
M S
MS
Iurkova
S M
SM
Andronova
A N
AN
Fedorov
S E
SE
Severin
E S
ES
Severin
D000818:Animals; D002942:Circular Dichroism; D016503:Drug Delivery Systems; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002479:Inclusion Bodies / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020673:Protein Renaturation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
C097353:AFP protein, human; D055767:Immobilized Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins