title: Recombinant expression and purification of human placental growth factor 1 and specific camel heavy chain polyclonal antibody preparation.
abstract: Placental growth factor (PlGF) is a member of the vascular endothelial growth factor (VEGF) family. Unlike VEGF, PlGF is dispensable for normal cell development as well as playing various roles in pathological angiogenesis which occurs in tissue ischemia, inflammation, and malignancy. The PlGF-1 has been considered as a potential candidate for the diagnosis and targeting of pathological angiogenesis. Camelidae serum contains an important fraction of functional antibodies, called heavy-chain antibodies (HcAbs) that are naturally devoid of light chains. Camelid HcAbs recognize their cognate antigens by a single variable-domain, referred to as VHH or Nanobody. Here, we describe the expression and purification of recombinant human PlGF-1 (rhPlGF-1). This protein was subsequently used for the preparation of camel heavy chain polyclonal antibody against rhPlGF-1. The recombinant expression plasmid pET-26b-hPlGF-1 was introduced into Escherichia coli BL21 cells to express the rhPlGF-1 protein. Purified rhPlGF-1 was used to immunize camel, the specific reactivity of HcAb was determined with ELISA and western blot. Western blot analysis indicated that the antiserum specifically reacted to the recombinant protein. The rhPlGF-1 protein and its antibody may be used for the development of detection assays needed for clinical research.
authors.affiliation: Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Roghaye
authors.initials: R
authors.lastname: Arezumand
authors.affiliation: Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Mahdian
authors.affiliation: Biotechnology Research Center, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Behdani
authors.affiliation: Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran ; Department of Genetics and Molecular Biology, Medical School of Isfahan University of Medical Science, Isfahan, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Khanahmad
authors.affiliation: Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Jahangir
authors.initials: J
authors.lastname: Langari
authors.affiliation: Department of Laboratory Animal Science, Research and Production Complex, Pasteur Institute of Iran, Karaj, Iran.
authors.forename: Nabiollah
authors.initials: N
authors.lastname: Namvarasl
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije University Brussel, Brussels, Belgium ; Department of Structural Biology, NSF, VIB, Brussels, Belgium.
authors.forename: Reza
authors.initials: R
authors.lastname: Hassanzadeh-Ghasabeh
authors.affiliation: Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Sirous
authors.initials: S
authors.lastname: Zeinali
keywords: Angiogenesis; Heavy chain antibody; PlGF; Polyclonal antibody; VHH