Aqueous extraction of recombinant human proinsulin from transgenic maize endosperm

Different plant species have been used as systems to produce recombinant proteins. Maize is a crop considered to have a large potential to produce high levels of recombinant proteins and is the host for the recombinant proteins from plants currently available on the market. In the development of a p...

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Bibliographic Details
Published in:Biotechnology progress 2005-09, Vol.21 (5), p.1466-1471
Main Authors: Farinas, C.S, Leite, A, Miranda, E.A
Format: Article
Language:English
Subjects:
aqueous solutions
Biological and medical sciences
Biotechnology
Chemical Fractionation - methods
Combinatorial Chemistry Techniques - methods
corn
endosperm
extraction
Fundamental and applied biological sciences. Psychology
gene expression
Humans
ionic strength
Plants, Genetically Modified - metabolism
proinsulin
Proinsulin - biosynthesis
Proinsulin - isolation & purification
Protein Engineering - methods
recombinant proteins
Seeds - metabolism
Solutions
transgenes
transgenic plants
Ultrafiltration - methods
Water - chemistry
Zea mays
Zea mays - genetics
Zea mays - metabolism
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Summary:Different plant species have been used as systems to produce recombinant proteins. Maize is a crop considered to have a large potential to produce high levels of recombinant proteins and is the host for the recombinant proteins from plants currently available on the market. In the development of a plant system to produce a recombinant proteins it is important to consider the costs related to downstream processing. Also, the steps necessary to achieve the protein purity required will be highly influenced by the quality of the extract obtained. In this study, we analyzed aqueous extracts from the endosperm of transgenic maize expressing recombinant human proinsulin (rhProinsulin). A study of the effects of the variables pH and ionic strength on the extraction efficiency was carried out using experimental design and response surface methodology. Besides the concentration of the recombinant protein, the characteristics of the extracts were evaluated in terms of concentration of native components (proteins, carbohydrates, and phenolic compounds) and extract filterability. The highest rhProinsulin concentration (97.33 ng/mL) was found with a 200 mM NaCl pH 10.0 extraction solution. Under this experimental condition the concentrations of total soluble proteins, carbohydrates, and phenolics were 2.01 mg/mL, 2.21 mg/mL, and 0.11 mmol/L, respectively.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp050103r