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A modified hybridoma technique for production of monoclonal antibodies having desired isotypes
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Document Title
A modified hybridoma technique for production of monoclonal antibodies having desired isotypes
Author
Apiratmateekul N, Phunpae P, Kasinrerk W
Name from Authors Collection
Affiliations
Chiang Mai University; Chiang Mai University; National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC)
Type
Article
Source Title
CYTOTECHNOLOGY
ISSN
0920-9069
Year
2009
Volume
60
Open Access
Green Published
Publisher
SPRINGER
DOI
10.1007/s10616-009-9213-0
Format
Abstract
In the present study, we describe a modified hybridoma technique for production of monoclonal antibodies (mAbs) having a desired isotype. Mice were immunized with the antigen of interest. After having reached a high antibody titer, cells expressing IgM or IgG molecules were isolated from spleen cells of the immunized mice using a Magnetic Cell Sorting System. The isolated cells were fused with myeloma cells using the conventional fusion protocol. With the isolated IgM(+) spleen cells, more than 75% (85 +/- A 7%; means +/- A SD) were IgM producing cells and a large number of IgM mAbs specific to the protein of interest were obtained. With the isolated IgG(+) spleen cells, 41 +/- A 40% of the generated hybridomas produced IgG antibody and no IgM producing hybridoma was generated. A large number of IgG mAbs specific to the protein of interest could be produced. The results indicate that the generated hybridomas produce corresponding antibody isotypes as expressed on the surface of their starting cells. The technique that we have developed will be very useful for production of desired mAbs having a specific isotype.
Funding Sponsor
National Center for Genetic Engineering and Biotechnology (BIOTEC) of the National Science and Technology Development Agency, Thailand; Thailand Research Fund; Ministry of Education, Thailand
Publication Source
WOS