Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells
Phytohemagglutinin-L (PHA-L) induced aggregation of SP2 myeloma and antibody-producing hybridoma cells. The aggregates were of diverse shapes, and major-axis length of the hybridoma aggregates was mostly 50-80 μm at 1 μg/ml PHA-L and 100-150 μm at 5 μg/ml. PHA-L did not suppress growth rate at these...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
1999
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| Online Access: | http://psasir.upm.edu.my/id/eprint/114072/ http://psasir.upm.edu.my/id/eprint/114072/1/114072.pdf |
| _version_ | 1848866396673409024 |
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| author | Takamatsu, Hiroyuki Kawajiri, Hiroo Takahashi, Yoshitaka Ali, Abdul Manaf |
| author_facet | Takamatsu, Hiroyuki Kawajiri, Hiroo Takahashi, Yoshitaka Ali, Abdul Manaf |
| author_sort | Takamatsu, Hiroyuki |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Phytohemagglutinin-L (PHA-L) induced aggregation of SP2 myeloma and antibody-producing hybridoma cells. The aggregates were of diverse shapes, and major-axis length of the hybridoma aggregates was mostly 50-80 μm at 1 μg/ml PHA-L and 100-150 μm at 5 μg/ml. PHA-L did not suppress growth rate at these concentrations. The aggregated cells had almost the same antibody productivity as that of non-aggregated cells in a static culture. Essentially, identical results were obtained with soybean agglutinin (SBA). On the other hand, pokeweed mitogen (PWM) did not induce apparent aggregation at a concentration of 10 μg/ml. These results suggest that lectin binding to particular carbohydrate moiety on the cell surface is necessary for cell agglutination. Using PHA-L, a 200-ml suspension culture of aggregated hybridoma cells was successfully performed in a spinner flask over 10 days. The aggregates were mainly globular with a diameter of 50-100 μm at 1-2 μg/ml PHA-L. The aggregation greatly enhanced settlement of hybridoma cells by gravity, and medium exchanges were thereby easily performed in a short period. During a course of the semi-continuous culture, antibody concentrations of culture medium were maintained at approximately 10 μg/ml which was comparable to that of static culture of the aggregated hybridoma cells. Thus, the lectin aggregation is applicable to the separation of culture medium from anchorage-independent cells like hybridomas, and can be employed in a large-scale commercial production of biologically active proteins. |
| first_indexed | 2025-11-15T14:19:56Z |
| format | Article |
| id | upm-114072 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T14:19:56Z |
| publishDate | 1999 |
| publisher | Elsevier |
| recordtype | eprints |
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| spelling | upm-1140722024-12-02T08:23:11Z http://psasir.upm.edu.my/id/eprint/114072/ Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells Takamatsu, Hiroyuki Kawajiri, Hiroo Takahashi, Yoshitaka Ali, Abdul Manaf Phytohemagglutinin-L (PHA-L) induced aggregation of SP2 myeloma and antibody-producing hybridoma cells. The aggregates were of diverse shapes, and major-axis length of the hybridoma aggregates was mostly 50-80 μm at 1 μg/ml PHA-L and 100-150 μm at 5 μg/ml. PHA-L did not suppress growth rate at these concentrations. The aggregated cells had almost the same antibody productivity as that of non-aggregated cells in a static culture. Essentially, identical results were obtained with soybean agglutinin (SBA). On the other hand, pokeweed mitogen (PWM) did not induce apparent aggregation at a concentration of 10 μg/ml. These results suggest that lectin binding to particular carbohydrate moiety on the cell surface is necessary for cell agglutination. Using PHA-L, a 200-ml suspension culture of aggregated hybridoma cells was successfully performed in a spinner flask over 10 days. The aggregates were mainly globular with a diameter of 50-100 μm at 1-2 μg/ml PHA-L. The aggregation greatly enhanced settlement of hybridoma cells by gravity, and medium exchanges were thereby easily performed in a short period. During a course of the semi-continuous culture, antibody concentrations of culture medium were maintained at approximately 10 μg/ml which was comparable to that of static culture of the aggregated hybridoma cells. Thus, the lectin aggregation is applicable to the separation of culture medium from anchorage-independent cells like hybridomas, and can be employed in a large-scale commercial production of biologically active proteins. Elsevier 1999 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/114072/1/114072.pdf Takamatsu, Hiroyuki and Kawajiri, Hiroo and Takahashi, Yoshitaka and Ali, Abdul Manaf (1999) Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells. Journal of Immunological Methods, 223 (2). pp. 165-170. ISSN 0022-1759; eISSN: 0022-1759 https://www.sciencedirect.com/science/article/pii/S0022175998002166?via%3Dihub 10.1016/s0022-1759(98)00216-6 |
| spellingShingle | Takamatsu, Hiroyuki Kawajiri, Hiroo Takahashi, Yoshitaka Ali, Abdul Manaf Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells |
| title | Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells |
| title_full | Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells |
| title_fullStr | Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells |
| title_full_unstemmed | Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells |
| title_short | Continuous antibody production by phytohemagglutinin-L-aggregated hybridoma cells |
| title_sort | continuous antibody production by phytohemagglutinin-l-aggregated hybridoma cells |
| url | http://psasir.upm.edu.my/id/eprint/114072/ http://psasir.upm.edu.my/id/eprint/114072/ http://psasir.upm.edu.my/id/eprint/114072/ http://psasir.upm.edu.my/id/eprint/114072/1/114072.pdf |