Construction and characterization of phage-displayed leukocyte surface molecule CD99

The phage display technique has been described for the production of various recombinant molecules. In the present report, we used this technique to display a leukocyte surface molecule, CD99. PCR subcloning of CD99 cDNA from the mammalian expression vector pCDM8 to the phagemid expression vector pC...

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Main Authors: Tayapiwatana C., Kasinrerk W.
Format: Article
Language:English
Published: 2014
Online Access:http://www.scopus.com/inward/record.url?eid=2-s2.0-0037210659&partnerID=40&md5=a418825a195b531a7f12f14fc2d8257f
http://www.ncbi.nlm.nih.gov/pubmed/12436316
http://cmuir.cmu.ac.th/handle/6653943832/850
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Institution: Chiang Mai University
Language: English
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spelling th-cmuir.6653943832-8502014-08-29T09:02:14Z Construction and characterization of phage-displayed leukocyte surface molecule CD99 Tayapiwatana C. Kasinrerk W. The phage display technique has been described for the production of various recombinant molecules. In the present report, we used this technique to display a leukocyte surface molecule, CD99. PCR subcloning of CD99 cDNA from the mammalian expression vector pCDM8 to the phagemid expression vector pComb3HSS was performed. The resulting phagemid, pComb3H-CD99, was transformed into Escherichia coli XL-1 Blue. CD99 was displayed on the phage particles following infection of the transformed E. coli with the filamentous phage VCSM13. Using sandwich ELISA, the filamentous phage-displayed CD99 was captured by a CD99 monoclonal antibody (mAb) then detected with anti-M13 conjugated to horseradish peroxidase, confirming that the CD99 molecule was displayed on the phage particles. The CD99-phages inhibited induction of Jurkat cell aggregation by CD99 mAb MT99/1. Proper folding of the displayed CD99 bioactive domain was inferred from this finding. Our results demonstrate that the phage display technique can be applied to the generation of full-length CD99 molecules. The phage carrying this cell surface protein will be useful for identification of its counter receptor or ligand. 2014-08-29T09:02:14Z 2014-08-29T09:02:14Z 2003 Article 01757598 10.1007/s00253-002-1146-x 12436316 AMBID http://www.scopus.com/inward/record.url?eid=2-s2.0-0037210659&partnerID=40&md5=a418825a195b531a7f12f14fc2d8257f http://www.ncbi.nlm.nih.gov/pubmed/12436316 http://cmuir.cmu.ac.th/handle/6653943832/850 English
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
language English
description The phage display technique has been described for the production of various recombinant molecules. In the present report, we used this technique to display a leukocyte surface molecule, CD99. PCR subcloning of CD99 cDNA from the mammalian expression vector pCDM8 to the phagemid expression vector pComb3HSS was performed. The resulting phagemid, pComb3H-CD99, was transformed into Escherichia coli XL-1 Blue. CD99 was displayed on the phage particles following infection of the transformed E. coli with the filamentous phage VCSM13. Using sandwich ELISA, the filamentous phage-displayed CD99 was captured by a CD99 monoclonal antibody (mAb) then detected with anti-M13 conjugated to horseradish peroxidase, confirming that the CD99 molecule was displayed on the phage particles. The CD99-phages inhibited induction of Jurkat cell aggregation by CD99 mAb MT99/1. Proper folding of the displayed CD99 bioactive domain was inferred from this finding. Our results demonstrate that the phage display technique can be applied to the generation of full-length CD99 molecules. The phage carrying this cell surface protein will be useful for identification of its counter receptor or ligand.
format Article
author Tayapiwatana C.
Kasinrerk W.
spellingShingle Tayapiwatana C.
Kasinrerk W.
Construction and characterization of phage-displayed leukocyte surface molecule CD99
author_facet Tayapiwatana C.
Kasinrerk W.
author_sort Tayapiwatana C.
title Construction and characterization of phage-displayed leukocyte surface molecule CD99
title_short Construction and characterization of phage-displayed leukocyte surface molecule CD99
title_full Construction and characterization of phage-displayed leukocyte surface molecule CD99
title_fullStr Construction and characterization of phage-displayed leukocyte surface molecule CD99
title_full_unstemmed Construction and characterization of phage-displayed leukocyte surface molecule CD99
title_sort construction and characterization of phage-displayed leukocyte surface molecule cd99
publishDate 2014
url http://www.scopus.com/inward/record.url?eid=2-s2.0-0037210659&partnerID=40&md5=a418825a195b531a7f12f14fc2d8257f
http://www.ncbi.nlm.nih.gov/pubmed/12436316
http://cmuir.cmu.ac.th/handle/6653943832/850
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