Recombinant expression and purification of human placental growth factor 1 and specific camel heavy chain polyclonal antibody preparation

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• 作者：Roghaye Arezum ; Reza Mahdian ; Mahdi Behdani ; Hossein Khanahmad ; Jahangir Langari ; Nabiollah Namvarasl ; Reza Hassanzadeh-Ghasabeh ; Sirous Zeinali
• 关键词：PlGF ; VHH ; Polyclonal antibody ; Angiogenesis ; Heavy chain antibody
• 刊名：Saudi Journal of Biological Sciences
• 出版年：January, 2014
• 年：2014
• 卷：21
• 期：1
• 页码：35-39
• 全文大小：978 K

文摘

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.