A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production

One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B str...

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Main Authors:	Tayapiwatana C., Chotpadiwetkul R., Kasinrerk W.
Format:	Article
Language:	English
Published:	2014
Online Access:	http://www.ncbi.nlm.nih.gov/pubmed/3502482 http://cmuir.cmu.ac.th/handle/6653943832/811
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Institution:	Chiang Mai University
Language:	English

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spelling	th-cmuir.6653943832-8112014-08-29T09:02:09Z A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production Tayapiwatana C. Chotpadiwetkul R. Kasinrerk W. One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B strain, survivin-biotin carboxyl carrier protein (BCCP) fusion protein was generated in the cytoplasm, where the BCCP domain serves as a target for in vivo biotinylation. The biotinylated heterologous protein was subsequently immobilized on streptavidin-coated magnetic particles and separated from other proteins in a magnetic field. The survivin-coated beads were used to raise immune responses in BALB/c mice for hybridoma production. A number of hybrid clones were found to secrete anti-survivin antibodies. Three established clones were selected for single cell cloning. All generated monoclonal antibodies specifically reacted with the standard human recombinant survivin. Two out of three monoclonal antibodies recognized survivin in tumor extracts. The present method has advantages in facilitating monoclonal antibody production by making antigen purification steps unnecessary. 2014-08-29T09:02:09Z 2014-08-29T09:02:09Z 2006 Journal Article 0022-1759 17109880 http://www.ncbi.nlm.nih.gov/pubmed/3502482 http://cmuir.cmu.ac.th/handle/6653943832/811 eng
institution	Chiang Mai University
building	Chiang Mai University Library
country	Thailand
collection	CMU Intellectual Repository
language	English
description	One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B strain, survivin-biotin carboxyl carrier protein (BCCP) fusion protein was generated in the cytoplasm, where the BCCP domain serves as a target for in vivo biotinylation. The biotinylated heterologous protein was subsequently immobilized on streptavidin-coated magnetic particles and separated from other proteins in a magnetic field. The survivin-coated beads were used to raise immune responses in BALB/c mice for hybridoma production. A number of hybrid clones were found to secrete anti-survivin antibodies. Three established clones were selected for single cell cloning. All generated monoclonal antibodies specifically reacted with the standard human recombinant survivin. Two out of three monoclonal antibodies recognized survivin in tumor extracts. The present method has advantages in facilitating monoclonal antibody production by making antigen purification steps unnecessary.
format	Article
author	Tayapiwatana C. Chotpadiwetkul R. Kasinrerk W.
spellingShingle	Tayapiwatana C. Chotpadiwetkul R. Kasinrerk W. A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
author_facet	Tayapiwatana C. Chotpadiwetkul R. Kasinrerk W.
author_sort	Tayapiwatana C.
title	A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
title_short	A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
title_full	A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
title_fullStr	A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
title_full_unstemmed	A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
title_sort	novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production
publishDate	2014
url	http://www.ncbi.nlm.nih.gov/pubmed/3502482 http://cmuir.cmu.ac.th/handle/6653943832/811
_version_	1681419553416937472

A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production

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