This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tachibana, H.
Right arrow Articles by Ihara, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tachibana, H.
Right arrow Articles by Ihara, S.

 Previous Article  |  Next Article 

Clinical and Diagnostic Laboratory Immunology, January 2004, p. 216-218, Vol. 11, No. 1
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.1.216-218.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Bacterial Expression of a Human Monoclonal Antibody-Alkaline Phosphatase Conjugate Specific for Entamoeba histolytica

Hiroshi Tachibana,1* Masataka Takekoshi,2 Xun-Jia Cheng,1 Yuta Nakata,1 Tsutomu Takeuchi,3 and Seiji Ihara2

Departments of Infectious Diseases,1 Molecular Life Sciences, Tokai University School of Medicine, Isehara, Kanagawa 259-1193,2 Department of Tropical Medicine and Parasitology, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan3

Received 1 August 2003/ Returned for modification 14 September 2003/ Accepted 2 October 2003

We previously produced human monoclonal antibody Fab fragments specific to Entamoeba histolytica in Escherichia coli. In order to use these Fab fragments for diagnostic purposes, an expression vector to produce a fusion protein of Fab and alkaline phosphatase (PhoA) in E. coli was designed and constructed. The E. coli PhoA gene was fused to the 3' terminus of the gene encoding the heavy-chain Fd region. The kappa and Fd genes from a previously prepared antibody clone, CP33, which is specific for the 260-kDa lectin of E. histolytica, were used as human antibody genes. When the fusion protein of CP33 and PhoA was incubated with paraformaldehyde-fixed trophozoites of E. histolytica and developed with a substrate, the trophozoites appeared to be stained. These results demonstrate the feasibility of bacterial expression of a human monoclonal antibody-PhoA conjugate specific for E. histolytica and that the antibody can be used to detect E. histolytica antigen without the use of chemically conjugated secondary antibodies.

* Corresponding author. Mailing address: Department of Infectious Diseases, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193, Japan. Phone: 81 (463) 93-1121. Fax: 81 (463) 95-5450. E-mail: htachiba{at}is.icc.u-tokai.ac.jp.

Clinical and Diagnostic Laboratory Immunology, January 2004, p. 216-218, Vol. 11, No. 1
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.1.216-218.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Tachibana, H., Matsumoto, N., Cheng, X.-J., Tsukamoto, H., Yoshihara, E. (2004). Improved Affinity of a Human Anti-Entamoeba histolytica Gal/GalNAc Lectin Fab Fragment by a Single Amino Acid Modification of the Light Chain. CVI 11: 1085-1088 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»