High-efficient expression, refolding and purification of functional recombinant C-terminal fragment of human alpha-fetoprotein.
Human alpha-fetoprotein (hAFP) is an oncofetal protein which is a common cancer marker. Conjugates of native hAFP with different cytostatic agents inhibit growth of cancer cells in vivo and in vitro. The hAFP interacts with its receptor (AFPR) on the surface of cancer cells via its C-terminal domain. The aim of this work was to develop a highly efficient expression system in Escherichia coli and efficient refolding procedure for the recombinant C-terminal fragment of hAFP (rAFP-Cterm) and to characterize its functional properties. C-terminal fragment of hAFP (rAFP-Cterm) comprising amino acids from 404 to 609 was expressed in E. coli BL21 (DE3) strain with high yield. High efficient purification and refolding procedures were developed giving yield of refolded protein about 80% with purity about 95%. The refolded rAFP-Cterm bound specifically with cancer cells carrying AFPR and was accumulated by them with the same efficiency as native hAFP. This rAFP-Cterm can be used as a vehicle for the targeted delivery of drugs to cancer cells.
Sharapova
Olga A
OA
Moscow Research Institute of Medical Ecology, Simpheropolski Blvd. 8, Moscow 117638, Russia. sharapova_o@hotmail.com
Pozdnykova
Natalia V
NV
Laurinavichyute
Daniela K
DK
Yurkova
Maria S
MS
Posypanova
Galina A
GA
Fedorov
Alexey N
AN
Severin
Sergey E
SE
Severin
Evgeny S
ES
D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D056148:Chromatography, Reverse-Phase; D002942:Circular Dichroism; D016503:Drug Delivery Systems; D006801:Humans; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011485:Protein Binding; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D000509:alpha-Fetoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
C097353:AFP protein, human; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins