Volume 16, Issue 3 (7-2012)                   IBJ 2012, 16(3): 126-121 | Back to browse issues page

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Habibi Anbouhi M, Feiz Barazandeh A, Bouzari S, Abolhassani M, Khanahmad H, Golkar M, et al . Functional Recombinant Extra Membrane Loop of Human CD20, an Alternative of the Full Length CD20 Antigen. IBJ 2012; 16 (3) :126-121
URL: http://ibj.pasteur.ac.ir/article-1-736-en.html
Background: 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 a polypeptide is easer and favorable comparing to that of a full length transmembrane protein CD20. Methods: 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. Results: 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. Conclusion: 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.
Keywords: E. Coli, CD20, Thioredoxin
Type of Study: Full Length | Subject: Related Fields

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