Non-mitogen containing conditioned medium for hybridoma production and single cell cloning

Background: The hybridoma technique is the standard method for production of monoclonal antibodies of interest. However, the newly formed hybridomas and cells at low density often grow poorly or die. This is the major obstacle to production of monoclonal antibodies. Objective: The aim of this study...

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Main Authors: Apiratmateekul,N., Pata,S., Chiampanichayakul,S., Kasinrerk,W.
Format: Article
Published: The Allergy and Immunology Society of Thailand 2015
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http://cmuir.cmu.ac.th/handle/6653943832/38104
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-381042015-06-16T07:38:24Z Non-mitogen containing conditioned medium for hybridoma production and single cell cloning Apiratmateekul,N. Pata,S. Chiampanichayakul,S. Kasinrerk,W. Immunology and Allergy Immunology Background: The hybridoma technique is the standard method for production of monoclonal antibodies of interest. However, the newly formed hybridomas and cells at low density often grow poorly or die. This is the major obstacle to production of monoclonal antibodies. Objective: The aim of this study was to establish a method for preparation of conditioned medium in the absence of mitogen for promoting the growth of hybridomas after cell fusion and during single cell cloning. Methods: Culture supernatants were obtained from the cultures of BW5147 mouse thymoma cells without mitogen stimulation. Novel conditioned mediums were investigated for their ability to support hybridoma single-cell cloning and hybridoma production. Results: We demonstrated that these conditioned mediums could support hybridoma single-cell growth, both for stable and newly generated hybridomas, at a level equal to the commercial conditioned medium BM-Condimed H1. The conditioned medium was most effective at a final concentration of 20% with the basal medium supplemented with 10% fetal calf serum. The novel conditioned medium could also be effectively employed for generation of hybridomas secreting various monoclonal antibodies. Interestingly, fibroblast overgrowth in the post-fusion wells was markedly reduced when using the novel conditioned medium, as compared to the commercial BM-Condimed H1, likely due to the absence of mitogen in the conditioned medium. Conclusion: We describe the method for production of a novel conditioned medium for hybridoma technology. This method is simple, needing no special technology or sophisticated equipment. The novel conditioned medium is, therefore, recommended for use in place of expensive commercial conditioned medium for hybridoma technology, especially in resource-limited countries. 2015-06-16T07:38:24Z 2015-06-16T07:38:24Z 2012-06-01 Article 0125877X 2-s2.0-84863469954 22830290 http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84863469954&origin=inward http://cmuir.cmu.ac.th/handle/6653943832/38104 The Allergy and Immunology Society of Thailand
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Immunology and Allergy
Immunology
spellingShingle Immunology and Allergy
Immunology
Apiratmateekul,N.
Pata,S.
Chiampanichayakul,S.
Kasinrerk,W.
Non-mitogen containing conditioned medium for hybridoma production and single cell cloning
description Background: The hybridoma technique is the standard method for production of monoclonal antibodies of interest. However, the newly formed hybridomas and cells at low density often grow poorly or die. This is the major obstacle to production of monoclonal antibodies. Objective: The aim of this study was to establish a method for preparation of conditioned medium in the absence of mitogen for promoting the growth of hybridomas after cell fusion and during single cell cloning. Methods: Culture supernatants were obtained from the cultures of BW5147 mouse thymoma cells without mitogen stimulation. Novel conditioned mediums were investigated for their ability to support hybridoma single-cell cloning and hybridoma production. Results: We demonstrated that these conditioned mediums could support hybridoma single-cell growth, both for stable and newly generated hybridomas, at a level equal to the commercial conditioned medium BM-Condimed H1. The conditioned medium was most effective at a final concentration of 20% with the basal medium supplemented with 10% fetal calf serum. The novel conditioned medium could also be effectively employed for generation of hybridomas secreting various monoclonal antibodies. Interestingly, fibroblast overgrowth in the post-fusion wells was markedly reduced when using the novel conditioned medium, as compared to the commercial BM-Condimed H1, likely due to the absence of mitogen in the conditioned medium. Conclusion: We describe the method for production of a novel conditioned medium for hybridoma technology. This method is simple, needing no special technology or sophisticated equipment. The novel conditioned medium is, therefore, recommended for use in place of expensive commercial conditioned medium for hybridoma technology, especially in resource-limited countries.
format Article
author Apiratmateekul,N.
Pata,S.
Chiampanichayakul,S.
Kasinrerk,W.
author_facet Apiratmateekul,N.
Pata,S.
Chiampanichayakul,S.
Kasinrerk,W.
author_sort Apiratmateekul,N.
title Non-mitogen containing conditioned medium for hybridoma production and single cell cloning
title_short Non-mitogen containing conditioned medium for hybridoma production and single cell cloning
title_full Non-mitogen containing conditioned medium for hybridoma production and single cell cloning
title_fullStr Non-mitogen containing conditioned medium for hybridoma production and single cell cloning
title_full_unstemmed Non-mitogen containing conditioned medium for hybridoma production and single cell cloning
title_sort non-mitogen containing conditioned medium for hybridoma production and single cell cloning
publisher The Allergy and Immunology Society of Thailand
publishDate 2015
url http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84863469954&origin=inward
http://cmuir.cmu.ac.th/handle/6653943832/38104
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