ORIGINAL RESEARCH PAPER Expression of human apolipoprotein A-I in Nicotiana tabacum Pasquale Chiaiese • Maria Minutolo • Angela Arciello • Fulvio Guglielmi • Renata Piccoli • Edgardo Filippone Received: 29 July 2010 / Accepted: 24 August 2010 / Published online: 5 September 2010 Ó Springer Science+Business Media B.V. 2010 Abstract Several transgenic tobacco lines express- ing human apolipoprotein A-I (ApoA-I) were obtained. Western blot analyses indicated the expression of the recombinant protein in plant organs at various stages of development, including senescent leaves. A cell line expressing human ApoA-I was established from a T 1 transgenic plant. Recombinant ApoA-I was isolated either from extracts of transgenic leaves and from the culture medium of transgenic cells using an antibody- based one-step procedure. Keywords Apolipoprotein A-I  Molecular farming  Nicotiana tabacum  Plant cell culture  Transgenic plant Introduction High density lipoproteins (HDL) play an important role in the catabolism of cholesterol by the ‘‘reverse cholesterol transport’’, a process that maintains con- stant the levels of free cholesterol in blood, preventing its pathogenic accumulation in the arteries. Apolipo- protein A-I (ApoA-I), the major HDL component (Dalton and Swaney 1993), is involved in the main steps of cholesterol transport. Human ApoA-I is synthesized in the liver and small intestine as a pre- pro-ApoA-I (Gordon et al. 1983). Upon cleavage of the 18-residue signal peptide (pre-peptide), the pro- protein is secreted into the plasma where it is converted to its mature form (Zannis et al. 1983). Recent studies on transgenic LDL receptor-defi- cient mice, lacking mouse ApoA-I and over-express- ing the human homolog, have shown that the lipoprotein has a strong protective action against atherosclerosis (Valenta et al. 2006). This finding suggests that an improvement in human lipoprotein profile by increasing ApoA-I levels reduces risks due to atherosclerosis. Purpose of work Apolipoprotein A-I (ApoA-I), the major component of plasma high-density lipoproteins, plays a pivotal role in preventing atherosclerosis development, being involved in the transport of cholesterol from peripheral cells to the liver where it is degraded. In this study, plants and plant cells cultures are suggested as suitable platforms for the production of ApoA-I for therapeutic or analytical purposes. Pasquale Chiaiese, Maria Minutolo contributed equally to this work. P. Chiaiese (&)  M. Minutolo  E. Filippone Department of Soil, Plant, Environmental and Animal Production Sciences, School of Biotechnological Sciences, University of Naples Federico II, Naples, Italy e-mail: chiaiese@unina.it A. Arciello  F. Guglielmi  R. Piccoli Department of Structural and Functional Biology, School of Biotechnological Sciences, University of Naples Federico II, Complesso Universitario di Monte S. Angelo Via Cinthia 4, 80126 Naples, Italy A. Arciello  R. Piccoli Istituto Nazionale di Biostrutture e Biosistemi (INBB), Rome, Italy 123 Biotechnol Lett (2011) 33:159–165 DOI 10.1007/s10529-010-0388-4