title: Functional recombinant extra membrane loop of human CD20, an alternative of the full length CD20 antigen.
abstract: Targeting of CD20 antigen with monoclonal antibodies has become the mainstay in the treatment of non-Hodgkin's lymphomas and immunotherapeutic depletion of malignant B cells. Accessibility of antigen is one of the crucial factors in development of monoclonal antibodies against this antigen. One major problem in expression of full length CD20 is aggregation and misfolding. Therefore, production of an alternative polypeptide is easer and favorable comparing to that of a full length transmembrane protein CD20.
  
  
  
  In this study, we expressed the extra membrane loop of hCD20 (exCD20) consisting of a non-glycosylated 47-amino acids region. The exCD20 coding sequence was amplified by PCR and cloned in pET32a(+) expression vector. The desired protein was expressed in fusion with thioredoxin and 6× His tag in E. coli Origami strain. ELISA and Western-blotting data were performed to indicate the functionality of this protein.
  
  
  
  We have obtained the exCD20 recombinant protein which can be detected in ELISA and Western-blot experiments. This recombinant fusion protein was soluble and stable without aggregation and misfolding problems.
  
  
  
  The recombinant extra membrane loop of human CD20 protein in fusion with thioredoxin (exCD20) can be used in function assays and some applications such as ELISA, immuneblotting, affinity purification, immunization, screening, and development of anti-CD20 antibodies.
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Habibi Anbouhi
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Aida
authors.initials: A
authors.lastname: Feiz Barazandeh
authors.affiliation: Molecular Biology Unit, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Saeid
authors.initials: S
authors.lastname: Bouzari
authors.affiliation: Hybridoma Laboratory, Dept. of Immunology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Abolhassani
authors.affiliation: Production and Research Complex, Pasteur Institute of Iran, Karaj, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Khanahmad
authors.affiliation: Molecular Parasitology Laboratory, Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Golkar
authors.affiliation: Dept. of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Aghasadeghi
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Behdani
authors.affiliation: Molecular Biology Unit, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Jahanian-Najafabadi
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Shokrgozar
mesh_terms: D000595:Amino Acid Sequence; D018951:Antigens, CD20 / Q000737:chemistry* / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry*; D003001:Cloning, Molecular; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D018951:Antigens, CD20; D011994:Recombinant Proteins