Process development and economic evaluation of recombinant human lactoferrin expressed in rice grain

Abstract

In this paper, we show that recombinant human lactoferrin (rhLF) has been stably expressed at 0.5% brown rice flour weight for nine generations. Process development indicates that rhLF can be efficiently extracted from rice flour in 20 mM phosphate buffer (pH 7.0) containing up to 0.5 M NaCl and at a ratio of 1 kg flour to 10 L buffer. After solid/liquid separation, the extract can then be loaded directly onto an ion-exchange column and rhLF can be eluted using 0.8 M NaCl. The resulting rhLF is about 95% pure. A range of biochemical and biophysical analyses were carried out and results indicated that the purified rhLF was identical to its native human counterpart other than its glycosylation. Economic analysis shows that at 600 kg/year scale, the cash cost to produce 1 g of rhLF of pharmaceutical grade is US$ 5.90. Analysis also indicates that the expression level has profound impact on costs related to planting, milling, extraction and purification, thus high level expression of recombinant protein in plants is one of the key parameters for the success of plant made pharmaceuticals.

Transgenic tobacco plants expressing antimicrobial peptide bovine lactoferricin show enhanced resistance to phytopathogens

“…A high concentration of lactoferrin is found in human milk and has been suggested to have several biological activities, including protection from pathogens, regulation of iron absorption, modulation of the immune system, and cellular growth promoting activity (Lönnerdal and Iyer 1995). Recombinant lactoferrin has been produced in plants to protect against plant pathogenic bacteria (Mitra and Zhang 1994;Zhang et al 1998) and to increase nutritive value (Nandi et al 2002;Nandi et al 2005). Lfc is the antimicrobial fragment derived from the full length lactoferrin protein upon pepsin cleavage.…”

Antibacterial Activity of Recombinant Human Lactoferrin from Rice: Effect of Heat Treatment

“…It was supplied in the apo form (0.05 mg of Fe/g of LF) and as-isolated from rice (0.98 mg of Fe/g of LF). The procedure for isolation and preparation of both lactoferrins has been described by Nandi et al 22) Human lactoferrin (hLF) was isolated from milk by the method of Bezwoda and Mansoor. 23) All lactoferrins were analysed by SDS-PAGE which showed a band corresponding to a protein with a molecular mass of about 80 kDa and a purity higher than 90%; therefore, they were used without further purification.…”

Interfacial assembly of proteins and peptides: recent examples studied by neutron reflection

“…The demand for LF has stimulated the development of various recombinant technologies for an efficient and economical production of LF. One successful product is the recombinant rice lactoferrin (rLF; Nandi et al 2005;Rachmawati et al 2005). The rLF has the same amino acid sequence and three-dimensional structure as hLF.…”