title: Recombinant human lactoferrin-Fc fusion with an improved plasma half-life.
abstract: Lactoferrin (LF), an 80-kDa iron-binding glycoprotein found in mammalian exocrine secretions, has potential therapeutic efficacy due to its extensive health-promoting effects. However, LF is rapidly cleared from the circulation (∼12.6min half-life for recombinant human LF [rhLF] in rats), which limits its therapeutic potential. Therefore, to improve plasma stability, we developed a recombinant human LF (hLF)-immunoglobulin G1 (IgG1) fragment crystallizable domain (Fc) fusion (hLF-hinge-CH2-CH3) expressed in a Chinese Hamster Ovary cell (CHO) expression system and evaluated the in vitro bioactivities and pharmacokinetic properties of the purified fusion. CHO DG44 cells were transfected with an expression vector coding for recombinant hLF-hinge-CH2-CH3. Iron binding, Caco-2 uptake, and thermal stability were investigated in vitro, and pharmacokinetic parameters were investigated in vivo. hLF-hinge-CH2-CH3 was significantly expressed in CHO cells (∼100mg/l culture), was readily purified, and exhibited 98.3% of the non-fused rhLF iron-binding activity. Caco-2 uptake and thermal stability were improved for hLF-Fc fusion relative to rhLF. Moreover, hLF-hinge-CH2-CH3 demonstrated a plasma half-life that was 9.1-fold longer than that of rhLF as well as longer than that of the PEGylated bovine LFs that we previously developed. Thus, CHO-derived hLF-hinge-CH2-CH3, with enhanced pharmacokinetic properties, is a promising candidate drug for potential parenteral administration.
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Shiga
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Oshima
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kojima
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Akinori
authors.initials: A
authors.lastname: Sugimoto
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Tamaki
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Murata
authors.affiliation: Department of Veterinary Medicine, Tottori University, 4-101 Koyama-Minami, Tottori 680-8553, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Takeuchi
authors.affiliation: School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan. Electronic address: atsato@stf.teu.ac.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sato
mesh_terms: D000818:Animals; D016466:CHO Cells; D018938:Caco-2 Cells; D003412:Cricetulus; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000097:blood / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D007074:Immunoglobulin G / Q000097:blood / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D007408:Intestinal Absorption; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000097:blood / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics; D008297:Male; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics
chemical_list: D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D007501:Iron; D007781:Lactoferrin
keywords: Fc fusion protein; Lactoferrin; Pharmacokinetics