Three-Dimensional Culture for Monoclonal Antibody Production by Hybridoma Cells Immobilized in Macroporous Gel Particles
(2008) In Biotechnology Progress 24(5). p.1122-1131- Abstract
- Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on... (More)
- Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAX (TM) ) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on Gluta- MAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mu g mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1283665
- author
- Nilsang, Suthasinee LU ; Nehru, Vishal LU ; Plieva, Fatima LU ; Kutty Selva, Nandakumar LU ; Rakshit, Sudip Kumar ; Holmdahl, Rikard LU ; Mattiasson, Bo LU and Kumar, Ashok LU
- organization
- publishing date
- 2008
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- macroporous gel particles, antibody production, M2139 hybridoma cells, 3D cultivation, medium optimization, cell immobilization
- in
- Biotechnology Progress
- volume
- 24
- issue
- 5
- pages
- 1122 - 1131
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000260738900015
- pmid:19194922
- scopus:56249132172
- ISSN
- 1520-6033
- DOI
- 10.1002/btpr.28
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Medical Inflammation Research (013212019), Biotechnology (LTH) (011001037)
- id
- 5f44a9ac-e6df-494c-8d7f-84601d214f11 (old id 1283665)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/19194922?dopt=Abstract
- date added to LUP
- 2016-04-01 12:35:02
- date last changed
- 2023-08-15 13:30:56
@article{5f44a9ac-e6df-494c-8d7f-84601d214f11,
abstract = {{Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAX (TM) ) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on Gluta- MAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mu g mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production.}},
author = {{Nilsang, Suthasinee and Nehru, Vishal and Plieva, Fatima and Kutty Selva, Nandakumar and Rakshit, Sudip Kumar and Holmdahl, Rikard and Mattiasson, Bo and Kumar, Ashok}},
issn = {{1520-6033}},
keywords = {{macroporous gel particles; antibody production; M2139 hybridoma cells; 3D cultivation; medium optimization; cell immobilization}},
language = {{eng}},
number = {{5}},
pages = {{1122--1131}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Biotechnology Progress}},
title = {{Three-Dimensional Culture for Monoclonal Antibody Production by Hybridoma Cells Immobilized in Macroporous Gel Particles}},
url = {{http://dx.doi.org/10.1002/btpr.28}},
doi = {{10.1002/btpr.28}},
volume = {{24}},
year = {{2008}},
}