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Abstract

Currently, Graft-versus-Host Disease (GvHD) has been the major barrier to the application of allogeneic bone marrow transplantation for hematopoietic disorders treatment, especially leukemia. GvHD is caused by donor mature T cells’ attack on recipient’s tissues and organs. Recently, T cell depletion using immunomagnetic nanoparticles is one of the most effective methods to prevent GvHD. In this present study, polyclonal antibodies against Jurkat T cell membrane were generated as a component of immunomagnetic particle. Firstly, Jurkat T cell membrane was fractionated by cloud point extraction using Triton X-114. Subsequently, the fractionated membrane was used to subcutaneously immunize rabbits for polyclonal antibodies production with a dose of 50 μg for priming and 25 μg for the next four boostings. Rabbit serum was collected and partially precipitated in 50 percent of saturated ammonium sulfate solution. Next, precipitated polyclonal antibodies were purified by using protein-A affinity chromatography column and the purity was determimed by SDS-PAGE. Afterwards, the purified antibodies were used in immune-fluorescent stainings and the recognition to Jurkat T cells was evaluated via fluorescent microscopic imaging. Finally, the purified antibodies were negatively selected by incubating with TF-1 cell, a hematopoietic stem cell, to eliminate cross-reacting antibodies. The immunocytofluorescent staining results showed that the purified and selected polyclonal antibodies weakly cross-reacted with TF-1 cells and strongly bound to Jurkat T cell



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Article Details

Issue: Vol 19 No 1 (2016)
Page No.: 26-35
Published: Mar 31, 2016
Section: Natural Sciences - Research article
DOI: https://doi.org/10.32508/stdj.v19i1.541

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Creative Commons License

Copyright: The Authors. This is an open access article distributed under the terms of the Creative Commons Attribution License CC-BY 4.0., which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 How to Cite
Trinh, T., Huynh, M., & Tran, H. (2016). Generation and selective isolation of IgG antibodies against Jurkat T-cell membrane proteins. Science and Technology Development Journal, 19(1), 26-35. https://doi.org/https://doi.org/10.32508/stdj.v19i1.541

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