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 [11–13]. The hAFP interacts with its receptor (AFPR) on the surface of cancer cells via its C-termina...

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Bibliographic Details
Published in:Protein expression and purification 2010-09, Vol.73 (1), p.31-35
Main Authors: Sharapova, Olga A., Pozdnykova, Natalia V., Laurinavichyute, Daniela K., Yurkova, Maria S., Posypanova, Galina A., Fedorov, Alexey N., Severin, Sergey E., Severin, Evgeny S.
Format: Article
Language:English
Subjects:
AFP
Alpha-fetoprotein
alpha-Fetoproteins - biosynthesis
alpha-Fetoproteins - chemistry
alpha-Fetoproteins - genetics
alpha-Fetoproteins - isolation & purification
Cell Line, Tumor
Chromatography, Gel
Chromatography, Reverse-Phase
Circular Dichroism
Drug Delivery Systems
Humans
Oncofetal protein
Peptide Fragments - biosynthesis
Peptide Fragments - chemistry
Peptide Fragments - genetics
Peptide Fragments - isolation & purification
Protein Binding
Protein Folding
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Refolding
Targeted drug delivery
Tumor
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Summary: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 [11–13]. 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.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2010.03.025